Inflammatory bowel diseases (IBDs) are related to nuclear factor erythroid 2-related factor 2 (Nrf2) dysregulation. In vitro and in vivo studies using phytocompounds as modulators of the Nrf2 signaling in IBD have already been published. However, no existing review emphasizes the whole scenario for the potential of plants and phytocompounds as regulators of Nrf2 in IBD models and colitis-associated colorectal carcinogenesis. For these reasons, this study aimed to build a review that could fill this void. The PubMed, EMBASE, COCHRANE, and Google Scholar databases were searched. The literature review showed that medicinal plants and phytochemicals regulated the Nrf2 on IBD and IBD-associated colorectal cancer by amplifying the expression of the Nrf2-mediated phase II detoxifying enzymes and diminishing NF-κB-related inflammation. These effects improve the bowel environment, mucosal barrier, colon, and crypt disruption, reduce ulceration and microbial translocation, and consequently, reduce the disease activity index (DAI). Moreover, the modulation of Nrf2 can regulate various genes involved in cellular redox, protein degradation, DNA repair, xenobiotic metabolism, and apoptosis, contributing to the prevention of colorectal cancer. For thousands of years, several plants have been used as the primary source of medicine. Many current drugs for typical disease treatment have originated from medicinal plants, such as metformin, used against diabetes, even though new drugs have been developed using synthetic chemistry. Plant and plant-derived phytochemicals have been considered due to their therapeutic applications, fewer side effects, and low costs in several conditions, such as inflammatory and immunomodulated diseases ,2,3,4. CTherapeutically, IBDs involve incurable diseases, and their treatments focus on inducing and maintaining remission periods, reducing the necessity of patients’ hospitalization and surgery. IBDs are recurrent and remissive gastrointestinal illnesses affecting approximately 0.3% of Western populations, with a peak onset between the ages of 20 and 30. Crohn’s disease (CD) and ulcerative colitis (UC) are the most typical representatives. Although IBD patients usually have gastrointestinal symptoms, CD and UC can be distinguished from each other using different pathological features, including histopathological analyses and physical and colonoscopy examinations. Signs and symptoms of systemic inflammation may also occur, resulting in extraintestinal manifestations ,6,7.Pathologically, the causes of IBDs are still not completely understood. Many authors highlight numerous genetic and environmental factors, including more than 100 genes and smoking, inadequate nutrition, or stress ,9. AmongThe unphysiological and sustained chronic inflammation environment promoted by IBD intestinal mucosal inflammation is also the major risk factor for the development of colitis-associated colorectal cancer. Colorectal-associated colitis carcinogenesis is recognized as the third most deadly cancer worldwide due to its high prevalence among the elderly population. It shares many molecular similarities with sporadic colorectal cancer, and preclinical investigators have demonstrated the potential role of Nrf2/Nuclear Factor kB (NF-κB) alterations even in the first stages of cancer occurrence ,13,14.Many different plants and phytochemicals have long been recognized to ameliorate inflammation and OS through Nrf2 signaling activation . AccordiCD is a severe chronic inflammatory disease of the gastrointestinal tract (GIT) with relapsing–remitting behavior. It is also called regional ileitis due to its frequent involvement of the ileum, which can occur anywhere in the GIT, being typically transmural. Its etiology is still unknown, but studies defend a multifactorial environmental, immunological, and genetic interaction that contributes to uncontrolled mucosal inflammation. The pathogenesis of CD involves changes in the intestinal microbiota, dysfunctions in the epithelium, and immune hyperreactivity, generating an uncontrolled inflammatory state due to dysregulation of the intestinal epithelial immune response, with the stimulation of pro-inflammatory and inhibition of regulatory pathways. Genetic factors play an essential role, as the concordance rate for monozygotic twins reaches 50%, contributing to the disease’s phenotypic expression. There are more than 160 genes associated with IBDs; many are shared between CD and UC. Several genes associated with IBD overlap with genes involved in the response against mycobacteria, supporting the idea that host interactions with such microorganisms are essential in the pathophysiology of CD. Environmental factors such as pathogenic microorganisms, microbiota, diet, smoking, and psychosocial factors also play an important role in its pathogenesis. Thus, improving life habits could change the microbiome to eubiosis, contributing to the stabilization of the disease ,87,88,89The genes involved in CD may be involved in three stages of inflammation, namely (1) the recognition of pathogens, (2) the elimination of pathogens via innate and cellular immunity, and (3) the prevention of the invasion of pathogens through the intestinal barrier. Therefore, in comparisons of single nucleotide polymorphisms (SNPs) in patients with and without IBDs, there are abnormalities mainly in the functions of modulators of immune function, autophagy, and epithelial function that participate in the interaction between the host and microorganism ,89.The gene most associated with CD is nucleotide oligomerization binding domain 2 (NOD2). It encodes an innate immune intracellular protein involved in recognizing bacteria, binding to bacterial peptidoglycans, and subsequent stimulation of the immune system, activating signaling cascades, including the NF-κB pathway, expressed mainly by immune cells and antigen-presenting cells (APCs). As a result, pro-inflammatory and antimicrobial molecules such as interleukin (IL)1-β, IL6, IL8, tumor factor necrosis alfa (TNF-α), and α-defensins are released, which recruit innate immune cells with subsequent activation of adaptive immunity. Furthermore, NOD2 is an essential regulator of cell proliferation, differentiation, and apoptosis mediated by the mitogen-activated protein kinase (MAPK) pathway. Mutations in this gene impair the recognition of invading pathogens by Paneth cells, causing inflammatory intestinal lesions and the lower release of antimicrobial proteins, such as α-defensin. These mutations are probably related to a weakened immune response due to lower activation of the NF-κB transcription factor. However, even with the increased risk associated with NOD2 polymorphisms, only 10% of people with such variants will develop the disease. Finally, NOD2 is central to bacterial autophagy, which initiates by recruiting the molecular autophagy ATG16L1 (type-16 related to autophagy) to the bacterial uptake site. The mutation in ATG16L1 increases the risk of developing IBDs by impairing the ability of autophagy in Paneth cells in cases of intracellular pathogen degradation, indirectly affecting their ability to secrete antimicrobial products, which favors bacterial proliferation and invasion. The ER stress of specialized intestinal epithelial cells (IECs) can be induced by various external factors, initiate a pathological unfolded protein response (UPR), and thus trigger intestinal inflammation. ATG16L1 polymorphisms in IECs determine the tolerable level of ER stress. It defines the activation of the UPR sensor inositol-requiring enzyme 1α (IRE1α), and the hyperactivation of this enzyme spontaneously generates ileitis in mice. Furthermore, signal transducer and activator of transcription 3 (STAT3) and NF-κβ signaling in IECs are also relevant since defects in these two pathways favor the formation of colitis. The NF-κβ kinase-α inhibitor (IKKα) phosphorylates ATG16L1 and thus prevents its degradation. Without this inhibitor, IRE1α accumulates and relays a pathological UPR. The IRGM gene (immunity-related GTPase M), which encodes guanosine triphosphate M, in addition to LRRK2, which encodes leucine-rich repeat protein kinase 2, when mutated, increases the risk of CD. Patients with CD with LRRK2 mutation have increased activation of intestinal dendritic cells, with the exaggerated release of pro-inflammatory molecules. The non-receptor tyrosine phosphatase type 2 (PTPN2) is an autophagy-related gene expressed by lymphoid and Paneth cells and causes the defective formation of autophagosomes. Its loss-of-function mutation in T cells increases the differentiation of T helper (TH) TH0 cells into TH1 and TH17 and reduces regulatory T cells. Therefore, the genes above manifest their polymorphisms as abnormalities in the activity of Paneth cells, which are IECs at the base of Lieberkühn’s crypts in the small intestine. The Cadherin 1 (CDH1) gene is susceptible to IBDs, thus increasing risks for CD. Its function encodes the epithelial protein E-cadherin, which is necessary for tight intestinal junctions. Mutations in this gene generate a more permeable intestinal wall, facilitating the invasion of pathogenic microbes. The HNF4-α gene encodes the hepatocyte nuclear factor 4-alpha, which modulates the expression of genes related to epithelial proliferation and tight junction formation, in addition to regulating adhesion and junction proteins, and, in its absence, malformation of the intestinal wall occurs. Human leukocyte antigen (HLA) genes play a role in the presentation of antigens to effector T cells and may be related to the acquisition of immunological tolerance. Specific mutations in its structure can generate greater risk, while others can act as protectors for CD ,86,87,89Several factors reinforce the role of mucosal immune responses in the pathogenesis of CD. Innate and adaptive immunities are the primary controllers of CD pathogenesis. The inflammatory process seems to be caused mainly by the failure of innate recognition of several bacteria, by the impaired intestinal barrier integrity, and by the spread of luminal pathogens through the intestinal wall, as previously mentioned, overloading innate immune cells to recognize and eliminate invaders ,84,89.Escherichia coli compared to patients with UC. Macrophages in a healthy intestinal mucosa control tissue remodeling by clearing apoptotic or senescent cells, and the alteration in their phagocytic function may contribute to the pathogenesis of CD. Studies have shown that such cells were significantly affected in CD patients, with reduced secretion of protective substances, such as IL-10, and increased synthesis of inflammatory molecules, generating a late elimination of microbial agents with an exacerbated inflammatory response. On the other hand, ILCs produce cytokines responsible for communication between the innate and adaptive immune systems. In patients with CD, in the inflamed ileum and colon, there is an increase in ILC1 and ILC3. Furthermore, areas with inflammation have high levels of interferon (IFN)-γ-producing ILC1s at the expense of ILC3, which synthesizes the anti-inflammatory cytokines IL-17 and IL-22, which suggests the increased transformation of ILC3 to ILC1, mediated by the inflammatory cytokine IL-12, may contribute to the disease [Cells such as neutrophils, macrophages, innate lymphoid cells (ILCs), monocytes, dendritic cells, and Paneth cells make up the intestinal innate immune response. Neutrophils, in a balanced environment, phagocytose pathogenic microorganisms. Still, when there is accumulation in the intestinal epithelium, it ends up compromising the intestinal epithelial barrier, stimulating the production of substances that result in inflammation. By phagocytosing microorganisms, they mainly fight extracellular bacteria and fungi, in addition to forming extracellular neutrophil traps (NETs), which immobilize large pathogens and allow localized lysis. Due to the digestive action of neutrophils, the damage may even be greater than the initial insult for which it had been recruited. In addition, the antimicrobial defense of neutrophils is defective in patients with CD, with impaired nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity of these cells and reduced inflammatory response against dead  disease ,89.Monocytes perform phagocytosis, respond to damage-associated molecular patterns (DAMPs) through toll-like receptor (TLR) signals, are tumor factor necrosis (TNF) producers, and stimulate mucosal inflammation, increasing tissue damage. In acute inflammation, monocytes differentiate into mature macrophages. In homeostasis, IECs produce transforming growth factor beta (TGF-β), thus increasing the synthesis of the anti-inflammatory cytokine IL-10 by dendritic cells. In inflammatory states, these cells express toll-like receptor 2 (TLR2) and TLR4, which stimulate the production of pro-inflammatory cytokines in scenarios of frequent interaction with the intestinal microbiota. They also promote cell activation and differentiation of TH0 cells into TH1, TH2, and TH17 and induce other essential inflammatory cells, such as natural killer cells. TH cells are activated and exert a TH1-like response. Most lymphocytes are activated in the intestinal lymphoid tissue and then go to the site of inflammation. Integrins, present on the leukocyte surface, are essential for leukocyte extravasation, as they make them bind to cell adhesion molecules present on the endothelial wall. In patients with CD, it is relevant that mucosal effector T cells may be resistant or less responsive to suppression mediated by Treg cells, which contributes to the maintenance of intestinal inflammation ,87,89,90Saccharomyces disruption antibodies (ASCAs). However, the contribution of plasma cells and B cells in the pathogenesis of IBDs has not yet been elucidated [Furthermore, humoral immunity is also relevant, given that altered levels of mucosal and secretory antibodies were detected in patients with CD with considerably elevated IgG levels against bacterial cytoplasmic proteins and reduced secretory IgA. In addition, CD patients often have anti-ucidated ,89,90.Thus, in a few words, CD originates due to the exacerbated responses of TH1 and TH17 cells to inflammatory cytokines synthesized by APCs and the release of pro-inflammatory cytokines IL-17, TNF, and IFN-γ, which maintain inflammation by stimulating the synthesis of more inflammatory cytokines, such as IL-1, IL-6, IL-8, IL-12, IL-18, and TNF by other cell types such as endothelial, monocytes, and macrophages. IL-12 and IL-23, produced by innate immune cells such as dendritic cells and macrophages and act as modulators of intestinal inflammation. Given the importance of inflammation in CD, current therapies block the synthesis of inflammatory mediators, stopping the accelerated inflammation ,89.Clostridium difficile, must be ruled out at the time of diagnosis. Nonetheless, the gold standard for diagnosis is ileum colonoscopy with biopsies [UC is defined as a continuous and diffuse inflammatory process that affects the epithelium and intestinal mucosa only of the colon and rectum. The classical clinical presentation is characterized by bloody diarrhea, abdominal pain, fecal urgency, and/or tenesmus. The prevalence of UC increases annually, ranging from 8.8 to 23.1 per 100,000 individuals-year. In North America, the range is 0.6 to 24.3 per 100,000 individuals-year, and in Europe, the range is 7.3 to 17.4 individuals-year. The disease has a principal peak in people in the second and third decades of life, regardless of sex, and a second smaller peak in the sixth and seventh decades of life ,92,93,94biopsies ,96.The objective of treatment is based on mucosal healing, and this is performed using drugs such as mesalamine in uncomplicated patterns of disease, glucocorticoids in acute cases, and, in more complicated forms, azathioprine, biological agents, Janus Kinases (JAK) inhibitors  and calcineurin inhibitors. Proctocolectomy should be considered in refractory cases or high-grade epithelial dysplasia ,97,98.Environmental and host factors influence the susceptibility to the development of UC. Antibiotic use, smoking, air pollution, mental and psychological state, physical exercise, and diet are fundamental points in the pathogenesis of IBDs. The pathology of the disease may start with events that impair the mucosal barrier, cause intestinal microbiota imbalance, and encourage an abnormal intestinal immune response. In addition, genetic predisposition is relevant for the development of the disease ,100.Although only 8% to 14% of patients with UC have a family history of IBD, first-degree relatives of people diagnosed with this condition have a 4-fold risk, monozygotic twins have concordance rates of 6% to 13%, and the Ashkenazi Jewish population, when compared to other ethnicities, has an increased risk of 3 to 5 times. It is known that multiple genetic polymorphisms drive the genetic basis of the disease and that genetic studies have found about 260 IBD-associated susceptibility loci . Many ofThe GIT mucosa is exposed to millions of antigens from food, the environment, and the microbiome. The GIT epithelium is covered with a thick layer of mucin , which provides a physical separation between antigens and intestinal immune cells but also has antimicrobial properties. In UC, the synthesis and secretion of mucin are impaired, which generates epithelial damage and increases the permeability of the mucosa to luminal pathogens, resulting in increased uptake of these antigens and increased potential for stimulation of the system. Concomitantly, it is believed that colonocytes, colonic epithelial cells, in UC have impaired expression of the peroxisome proliferator-activated receptor g (PPAR g), a nuclear receptor that regulates inflammation negatively .Activation of the innate immune response by antigens occurs through antigen-presenting cells and T cells. In addition, there is also an increase in the activation and sensitivity of mature dendritic cells, helping to generate inflammation. These dendritic cells express an abundance of TLR, which use pathogen pattern recognition to signal the activation of multiple transcription factors, such as the NF-κB transcription factor, which triggers inflammatory cascades. These inflammatory cascades produce pro-inflammatory cytokines, such as TNF-α, IL-12, IL-23, IL-21, and IL-24. These cytokines send signals through intracellular proteins, such as JAK, which further potentiate the activation and proliferation of lymphocytes .Another cell involved in the immune response is neutrophils, which are recruited in abundance with a histology characteristic of “crypt abscesses” in UC. The inflammatory environment of UC promotes neutrophil survival ). This increased survival increases neutrophils’ inflammatory actions and the consequent tissue damage through reactive oxygen species (ROS) and pro-inflammatory cytokines production. Furthermore, the formation of NETs functions as a reservoir for immunogenic molecules that sustain the inflammatory response. Therefore, it is believed that after disease onset, the previous wave of innate inflammatory neutrophils and monocytes  creates an inflammatory environment. The adaptive immune response of UC occurs after the initial presentation of antigens to CD4 T cells, in which they promote a non-classical Th2 response, with augmented levels of IL-4, IL-5, and IL-13 but mainly IL-4 .The general notion of Nrf2 emerged in the 1970s when investigations into the transcriptional control of its antioxidant response began. From this, several studies started on its anticancer effects due to its ability to increase detoxifying phase II and activating enzymes such as glutathione-S-transferase ,106.In 1989, the action of activator protein 1 (AP-1) and erythroid nuclear factor 2 (NF-E2) was described, the latter being identified as the first of the six cap’n’ transcription factors collar, with nuclear respiratory factor (NRF) 1, NRF2, NRF3, BTB and CNC homology (BACH) 1, and BACH2 identified shortly thereafter. The NF-E2/AP-1 sequence was associated with a promoter of the glutathione transferase (GST)-Ya gene, being responsive to t-butyl hydroquinone (t-BHQ), when it resulted in responses to its antioxidant action, giving rise to the term “antioxidant response elements” (AREs). This fact then indicated that the transcription of phase II detoxifying enzymes was sensitive and directly influenced by electrophiles, a key functional feature of most Nrf2 activators. With this idea, it was believed that the redox sensitivity of Nrf2 was conferred by a regulator capable of detecting oxidative/electrophilic stress ,109,110.The molecular interactions that act on Nrf2 signaling were uncovered in the 1990s when it was first determined that this molecule is downregulated by Keap1. In the 2000s, it was discovered that cysteines interacted with Keap1 in the electrophilic stress pathway and were modulated to different inducers, which became known as the “cysteine code’’ .Until now, Nrf2 was described as a molecule regulating inflammatory metabolism, expressed in several tissues and organs, such as the heart, brain, liver, kidney, and skin. Nrf2 signaling is gaining ground as it covers crucial roles in essential processes in the body, such as embryonic development, OS, and ischemia/reperfusion injury. In addition, there are reports that this molecule regulates free radical deputation and lipid homeostasis ,116,117./β-TrCP complex), which acts both in the nucleus and in the cytosol through metabolic changes; and, finally, the endoplasmic reticulum (ER)-transmembrane E3 ubiquitin ligase (HRD1), which is located in the endoplasmic reticulum and ubiquitylates Nrf2 during stress in this cellular compartment [Epigenetic factors and interaction with several signaling pathways work in the maintenance of Nrf2 levels. In addition to Keap1, the regulation also happens by three other ubiquitin ligase E3 complexes responsible for the ubiquitylation and degradation of Nrf2 including the Cullin 3 (CUL3)-RING-box protein 1 (RBX1)-KEAP1 complex, which is responsible for the response to electrophilic/OS in the cytosol; the Skp, Cullin, F-box containing complex  domains Neh1-Neh7, which help regulate and stabilize Nrf2 ,122. UndPost-transcriptional processing is involved as another activation mechanism, and the correct functioning of Nrf2 is modifications of the Nrf2 gene (NFE2L2) mRNA transcript. This event occurs from regulation by microRNAs (miRNAs), adenosine methylation, long non-coding RNAs (lncRNAs), and alternative splicing of the NFE2L2 transcript. Thus, miRNAs down-regulate or up-regulate the expression and activity of the Nrf2 protein by directly targeting the 3′-untranslated region (UTR) sequences of NFE2L2 or KEAP1 mRNA, respectively ,125. AftAs a consequence of this series of actions, several downstream antioxidants are activated, which generates positive feedback on the expression levels of inflammatory proteins and detoxification enzymes. The transcribed genes involve superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPX), and heme oxygenase (HO-1), which have been identified to suppress ROS and reduce OS ,126.There is growing evidence to suggest that Nrf2 is essential in OS injury, where Nrf2 is involved in several signaling pathways, including NF-κB and other cytokine signaling pathways . Thus, tAs Nrf2/Keap1 signaling plays a critical role in the maintenance of intracellular redox homeostasis, it also plays a role in a wide range of cells and organs in different types of conditions, such as cardiac diseases related to oxidative damage, including ischemic myocardial disease, heart failure and hypertrophic heart disease . In otheAs seen before, the regulation of Nrf2 depends on many factors, such as genetics, drugs, and the molecular environment. Phytochemicals have been documented as modulators of Nrf2 signaling, reversing hypermethylated states by inhibiting DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) by inducing translocation enzymes or even by inducing miRNA to target the 3′-UTR of the corresponding mRNA transcripts, which contains around 22 nucleotides ,132,135.Many compounds such as curcumin, sulforaphane, resveratrol, and 3,3′-diindolylmethane can protect cells from inflammation and OS by mediating the Nrf2 defense response and stimulating phase II detoxification enzymes, transporters, and antioxidants, which is effective in preventing further damage ,139,140.As aforementioned, Nrf2 corresponds to a stress-responsive transcription factor associated with maintaining cellular homeostasis and, under normal conditions, is kept in the cytoplasm of the cells by Keap1. The primary function of Keap1 is to facilitate the degradation of Nrf2 when it is not necessary. However, when OS achieves normal cells, these events trigger the dissociation between Keap1 and Nrf2, dissolving the Nrf2/Keap1 complex and promoting the translocation of Nrf2 to the cells’ nucleus where Nrf2 induces the activation and transcription of a variety of anti-oxidative and anti-inflammatory genes ,121,141.During the development of gastrointestinal diseases, the Nrf2/Keap1 axis plays a crucial role in maintaining bowel health and proper functionality. It is known that the Nrf2/Keap1 pathway is not only directly involved in the prevention of IBDs but also in the prevention of CD- and UC-related complications, such as intestinal fibrosis and colorectal cancer. Although the roles of Nrf2/Keap1 in the development of UC are still elusive, this complex’s functions in CD are better described. Most studies concentrate on the abilities of the complex to modulate the production of many pro-inflammatory cytokines , controlling the levels of antioxidant enzymes (such as HO-1 and NAD(P)H quinone oxidoreductase-1 (NQO1)) and extracellular matrix (ECM) degradation proteins ) and modulating intestinal cells’ autophagy . HoweverInflammation results in the production of ROS, which are cardinal events in the physiopathology of IBDs and lead to significant activation of Nrf2. Nrf2′s control of the inflammatory mediator production relies principally on the fact that Nrf2 possesses immunomodulatory functions, which affect the Nrf2-Keap1 pathway and protect host cells from inflammatory disorders. However, the tasks of the Nrf2-Keap1 axis are not always unidirectional for protection but depend on the type of immune cell targeted. As an example, the activation of Nrf2 in macrophages suppresses the activation of pro-inflammatory genes in these cells, which decreases the production of ROS and TNF-α, in addition to reducing the expression of TLR4 and the Forkhead box protein O1 (FOXO1) genes, which also decreases the production of IL-1β, IFN-β, and IL-6 and subsequent tissue injury. From another perspective, the disruption of Keap1 in myeloid cells was found to increase the bacterial phagocytic activity of peritoneal macrophages. On the other hand, Nrf2-deficient peritoneal neutrophils were found to increase the expression of IL-6, monocyte chemoattractant protein-1, TNF-α, and macrophage inflammatory protein-2 . AdditioThe Nrf2-Keap1 axis also causes effects in a cluster of differentiation 8 (CD8+) antigen-specific T cells. Sha et al.  reportedHowever, Nrf2 inhibits NF-κB activity via different mechanisms. During the development of IBDs, the activation of inflammatory pathways is fundamental and results in the production of many pro-inflammatory cytokines and other pro-inflammatory mediators, such as IFN-γ, TNF-α, IL-1β, and IL-12. These changes in IBDs are related mainly to the activation of the NF-κB inflammatory signaling associated with an exacerbated immunological response against the increased pathogenic bacteria concerning commensals through immune cells such as macrophages, monocytes, lymphocytes, and epithelial cells of the intestinal lymphoid tissue. NF-κB is central to inflammatory responses, and its expression depends on the redox status. When the levels of ROS and reactive nitrogen species (RNS) are elevated, these molecules stimulate and exacerbate the inflammation that is mechanistically linked to the p65 subunit of the NF-κB signaling pathway. During quiescence, NF-κB remains inactivated physically due to the direct interaction with its inhibitor, IkBα, which prevents the nuclear localization of NF-κB and its further stimulative actions on pro-inflammatory genes expression. However, during expressive redox disruption stimuli, IkB kinase (IKKβ) is activated and phosphorylates IkBα, which is then targeted for degradation. With the IkBα degradation, this inhibitor is removed from NF-κB and translates the factor to the cell’s nucleus. Once inside the nucleus and in contact with the gene expression machinery, NF-κB can modulate and exacerbate inflammation .The interplay between Nrf2 and NF-κB is a complex game in which many aspects of their relationship must be analyzed. Firstly, it is known that IKKβ possesses an ETGE motif that can bind Keap1 to target it for ubiquitination and further proteasomal degradation. Thus, under conditions of massive oxidation, such as during ROS abundance, Keap1 is inhibited, thereby signalizing to IKKβ, which in turn leads to the phosphorylation of IκBα and its further degradation. These events may result in an aberrant induction of NF-κB activation and, thus, in a massive expression of pro-inflammatory stimuli. Nevertheless, NF-κB transcriptionally activates at a specific kB promoter site the expression of Nrf2 signaling, thus tenting against its activation. This happens due to the competition between NF-κB and Nrf2 for the same coactivator histone acetyltransferase CREB-binding protein (CBP)/p300. In this competition, NF-κB, when up-regulated, may disrupt the interaction between the Nrf2 and the CBP’s CH1-KIX domains, thus decreasing Nrf2 activation .In addition, NF-κB also promotes the recruitment of many corepressor histone deacetylases 3 (HDAC3) in an attempt to induce the Nrf2-driven expression of ARE . HoweverSimilarly, Nrf2 inhibits the most important inflammatory factor in IBDs, the NF-κB, through several different cellular and biochemical mechanisms. Chronically, it is known that NF-κB activity is inhibited when Nrf2 expression is the most active. Nrf2 primarily inhibits NF-κB by suppressing ROS. Nrf2 activators reduce ROS presence and inhibit the ROS-mediated activation of the NF-κB-dependent production of pro-inflammatory factors . In concSpecifically, Nrf2 also beneficially affects cyclooxygenase (COX) and inducible nitric oxide (NO) synthase (iNOS) signaling in immunological and inflammatory cells. iNOS corresponds to a pro-inflammatory marker expressed mainly by immunological cells such as T cells, macrophages, and mature dendritic cells. COXs, especially COX-2, are critical enzymes in the inflammatory processes that catalyze the formation of prostaglandins and thromboxane during the arachidonic acid pathway. COX-2 is expressed mainly by macrophages and is up-regulated in response to various growth factors and other inflammatory stimuli, such as in the case of LPSs ,81. It wThe levels of proteases such as MMPs, which are zinc-dependent proteolytic enzymes responsible for the occurrence of the extracellular matrix components degradation, antiproteases, and OS parameters, are interconnected. They are also crucial in the Nrf2-mediated regulation of inflammatory processes . Nrf2 ovAs a transcription factor, Nrf2 regulates various genes involved in cellular redox homeostasis and protein degradation, DNA repair, and xenobiotic metabolism. Due to these reasons, the complex role of Nrf2 signaling in cancer evolves continuously since Nrf2 itself is responsible for the mechanisms that preserve cell function and viability during stress. However, the mode of Nrf2 activation often dictates its role in either preventing cancer development or promoting cancer progression. The wide array of cytoprotective target genes controlled by Nrf2 is fundamental in controlling and preventing oxidative, proteotoxic, and metabolic stresses that may contribute to cell malignant transformation. While Nrf2 activation plays a key role in chemoprevention, it was evidenced that prolonged Nrf2 activation can promote cancer initiation and progression as Nrf2 signaling is up-regulated during the first stages of tumor formation. Although this can be just a secondary effect of increased stress associated with the cell malignant transformation, whether Nrf2 is involved in cancer development remains unclear. However, it is known that prolonged Nrf2 activation associates highly with suppressed apoptosis of newly transformed cells by continuous up-regulation and activation of detoxification and DNA repairments. In addition, it stands to reason that the cell malignant transformation itself may not involve Nrf2. Still, the up-regulation of this signaling pathway is involved in mitigating the stress generated by replication errors, which helps mediate tumor growth and survival, as well as cancer initiation and also cancer progression ,163,164.4, compared with wild-type mice. Finally, as expected, the knockout mice did not present elevated phase II detoxifying/antioxidant enzymes NQO1 and UDP-glucuronosyltransferase 1A1.Colonic tumorigenesis occurs more frequently in males than in females due principally to lifestyle habits. In colon tumors, Nrf2 plays differential roles in the stages of tumorigenesis. Many studies regarding the purpose of investigating the roles of Nrf2 in colitis-associated tumorigenesis were published in the global scientific literature. Nonetheless, the majority of these studies are in vitro or in vivo and not clinical. C.-H. Song et al.  conducteCynara Cardunculus leaf extract against Nrf2 deactivation. The Caco-2 cells presented elevated NF-κB activation and ROS intracellular levels and decreased TAA and GSH levels. The plant extract was rich in chlorogenic acid and luteolin, and the results showed that the extract’s use could increase Nrf2 expression via increases in the Nrf2 nuclear levels, glutamate—cysteine ligase catalytic subunit (GCLC) mRNA, and NQO1 mRNA.Speciale, Muscarà, et al.  worked oPanax ginseng root extract rich in ginsenosides to evaluate its effectiveness in activating Nrf2 signaling to reduce inflammatory and oxidative stimulation in both an in vitro and an in vivo model. The models presented high levels of NO, TNF-α, IL-6, IL-1β, and ROS, in addition to augmented mitochondrial dysfunction. The results showed that the extract in different concentrations increased the Nrf2 nuclear translocation due to increased p62 phosphorylation and protein kinase b (Akt)-mammalian target of rapamycin (mTOR) signaling.S. Yang et al., 2022  worked wRose odorata sweet var. gigantean (Coll. Et Hemsl.) Rehd. Et Wils root extract (ROE) in activating Nrf2 signaling using a DSS-induced mice model of colitis. Due to the DSS administration, the mice presented elevations in the disease activity index (DAI), OS , and myeloperoxidase (MPO) levels), loss of epithelial and goblet cells, crypt aberrations, and prominent transmural inflammatory cells infiltration . After treatment with different ROE doses, the mice presented elevations in Nrf2 signaling due to principally increased HO-1 levels, as well as increased SOD and decreased numbers of nuclear NF-κB-p65 positive cells, p-NF-κB-p65, p-IKKα/β, and Keap1.Li et al., 2020  evaluateFicus pandurata Hance. could significantly ameliorate the disease process. The mice presented increased DAI and augmented intestinal inflammation and OS parameters through increased NF-κB activation and MDA levels and decreased intestinal barrier integrity, diamine oxidase activity (DAO), SOD, and glutathione peroxidase (GSH-Px). After the treatment, the mice presented elevated Nrf2 activation through diminished TLR4/ Innate Immune Signal Transduction Adaptor (MyD88) /NF-κB activation and Keap1, cytochrome b(558) subunit beta or NADPH (nicotinamide adenine dinucleotide phosphate hydrogen) oxidase 2 (NOX-2), and human neutrophil cytochrome b light chain (p22-phox) levels and augmented DAO, total SOD (T-SOD), HO-1, and NQO1 levels and Nrf2 translocation.Dai et al., 2021 [Moringa oleifera Lam. seed extract on the Nrf2 signaling pathway activation. The mice presented increased DAI and intestinal inflammation  and OS (increased MPO), as well as decreased intestinal integrity. During the experiment, Nrf2 signaling increased due to decreased pro-inflammatory gene expression, resulting in decreased NF-κB-dependent pro-inflammatory signals. Additionally, the mice presented elevated expression of the transcription factor STP1, NQO1, and HO-1.Kim et al., 2017 , during Aucklandia lappa Decne in colitis. The studied mice presented an elevated disease activity index (DAI). Both models expressed increases in TNF-α, IL-1β, and IL-6 levels, M1 macrophage polarization, iNOS, and COX-2 levels, NF-κB-related proteins expression, and p38 MAPK, NF-κB p65, extracellular signal-regulated kinases (ERK), and JNK phosphorylation, as well as decreased heme oxygenase 1 gene (HMOX-1) expression. After treatment, the models presented decreases in IL-1β mRNA, IL-6 mRNA, and TNF-α mRNA expression, MAPK, and NF-κB activation, and increased HMOX-1 mRNA, and ↑Nrf2 mRNA expression in the mice and decreased p38MAPK, p-NF-κB-p65, ERK, and JNK phosphorylation and increased Nrf2 and HMOX-1 expression in the cells.The in vivo and in vitro study by Chen et al., 2022 [Vaccinium myrtillus and Ribes nigrum extract, the cells presented diminished NF-κB-p65 translocation and overactivation of the Nrf2 pathways measured by increased Nrf2 nuclear translocation and NQO1 mRNA expression.Speciale, Bashllari, et al., 2022  conducteMesua assamica in NF-κB-RE-luc2P HEK 293 TNF-α-stimulated cells and a DSS-induced C57BL/6 mice model of colitis. The treated cells presented elevated intestinal inflammation and OS. In turn, the animals presented elevated DAI and splenomegaly, as well as reduced intestinal integrity and colon length. The models showed elevated levels of MPO, nitrite, IL-6, IL-1β, and TNF-α and reduced levels of GSH. After treatment with the plant, Nrf2 signaling was activated due to diminished NF-kB-related proteins expression, STAT3 signaling, and IKBα phosphorylation, as well as elevated SOD2 and HO-1 levels and NAD-dependent deacetylase sirtuin-1 (SIRT1) expression.Puppala et al., 2022  evaluateVaccinium myrtillus berry anthocyanin extracts in a DSS-induced Drosophila melanogaster model of intestinal inflammatory damage. Before treatment, the flies presented diminished intestinal integrity and length and elevated levels of ROS and pro-inflammatory cytokines. The signals of inflamed bowel disease disappeared due to Nrf2 activation, which was measured by high genetic expression of Nrf2-related proteins such as GCL, GSTS, and SOD.Zhang et al., 2022  evaluateVitis vinifera. Before treatment, the animals presented diminished body weight, elevated diarrhea and bloody stool, and elevated levels of pro-inflammatory cytokines , pro-oxidative enzymes, and NF-κB-related proteins expression, as well as decreased levels of HO-1 and SOD. After treatment, the mice started to present elevated HO-1 levels, elevated Nrf2-related protein gene expression, and decreased NF-κB-related protein expression.Wang et al.  worked wAcanthopanax senticosus against intestinal inflammation and colitis. The authors used hydrogen peroxide (H2O2)-induced RAW 264.7 cells and DSS-induced C57BL/6 mice to show that the activation of Nrf2 by the plant’s extract had a role in alleviating the disease. Before treatment, the models presented elevated inflammatory (elevated Keap1 levels), oxidative stimuli , SOD, GPx, and HO-1 levels), and elevated DAI scores in vivo. After treatment, the models presented decreased Keap1 and elevated HO-1 expression, which involves augmented nuclear translocation of the Nrf2.Su et al., 2022  evaluateForsythia suspensa extract, a well-known traditional medicine in Asian countries such as China, Japan, and Korea, against colitis induced by DSS in C57BL/6 mice. Before treatment with the extract, the mice presented elevated DAI, intestinal villi degeneration, intestinal necrosis, elevated proliferation and infiltration of inflammatory cells in the intestine, and diminished body weight. Additionally, the mice presented augmented levels of IL-1β, MDA, and MPO, as well as reduced levels of SOD. After treatment with the extract, the situation was reversed, and the mice started to present augmented Nrf2 activity measured by decreased levels of MDA, MPO, caspase-1, IL-1β, Gasdermin D, Keap1 activity and NLR family pyrin domain containing 3 (NLRP3) expression, ROS levels, pyroptosis, and ASC and by increased levels of SOD, HO-1, and NQO1.Chao et al.  studied Artemisia argyi ethanolic extract, also known as wormwood and Chinese mugwort, to evaluate its effects against the DSS-induced C57BL/6 mice model of colitis. The animals presented elevated DAI, colonic dysplasia, colon barrier disruption, and decreased body weight before treatment. Additionally, the mice presented elevated aspartate aminotransferase (AST), alanine aminotransferase (ALT), IL-6, IL-1β, TNF-α, and COX-2 levels. After treatment, the mice regulated Nrf2. Additionally, the animals presented decreased expression levels of IL-6, IL-1β, TNF-α, p-IκBα, p-NF-kB, COX-2 expression, intercellular adhesion molecule 1 (ICAM-1) expression, MCP1 expression, and iNOS expression, as well as elevated HO-1 levels.Shin et al.  used ArtDendrobium fimbriatum polysaccharide extract against this IBD sample. Before treatment, the mice presented elevated DAI and colon barrier disruption, decreased body weight, and augmented NF-κB signaling and Th17/regulatory T homeostasis disruption. After treatment, the mice presented diminished NF-κB signaling activation and Th17/regulatory T homeostasis disruption, as well as increased Nrf2 signaling activation.Wang et al.  conductePisum sativum L. hull polyphenol extracts in a DSS-induced C57BL/6 mice model of colitis. Before treatment, the animals presented high values of DAI and a high prevalence of diarrhea, blood in the stool, and reductions in colon length and body weight. The mice also presented augmented MPO, diminished claudin-1, occluding, and tight junction protein-1 (ZO-1), increased MDA, TNF-α, IL-1β, and IL-6 expressions, and decreased SOD, CAT, total antioxidant capacity (T-AOC), and IL-10 levels, as well as increased Keap1 and decreased Nrf2, GCLC, HO-1, and NQO1 expression levels. After treatment, the animals presented elevated Nrf2 expression and nuclear translocation, GCLC, HO-1, NQO1, IL-10, SOD, CAT, T-AOC, claudin-1, occludin, and ZO-1 expression, in addition to decreased MPO, MDA, TNF-α, IL-1β, IL-6, and Keap1 expressions.Guo et al.  studied Tetrastigma hemsleyanum leaf bio compounds and metabolites in a DSS-induced mice model of colitis. Before treatment, the animals presented diminished colon length and body weight, as well as reduced claudin-1, occludin, ZO-1, IL-10, Nrf2 nuclear translocation, SOD, CAT, HO-1, NQO1, and GCLC levels, in addition to augmented MPO, MDA, IL-1β, IL-6 and TNF-α levels. After treatment, the animals showed elevated Nrf2 nuclear translocation, increased claudin-1, occluding, ZO-1 expressions, IL-10, SOD, CAT, HO-1, NQO1, and GCLC expressions. The mice also presented reduced IL-1β, IL-6, TNF-α, and MPO expression.Wu et al.  studied Rhus chinensis Mill. fruit ethanolic extract. Before colitis induction, the mice presented elevated DAI, colon barrier disruption, and diminished colon length. Additionally, the animals presented elevated MPO, MDA, IL-1β, IL-6, TNF-α, p-NF-kB, p-IκB, COX-2, iNOS, p-P38, p-ERK1/2, and p-JNK levels, as well as reduced SOD, GSH, claudin-1, occludin, ZO-1, Nrf2 activation, NQO1, and HO-1. After treatment, the mice presented reduced MPO, MDA, IL-1β, IL-6, TNF-α, p-NF-kB, p-IκB, COX-2, iNOS, p-P38, p-Erk1/2, and p-JNK expression, as well as elevated SOD, GSH, claudin-1, occludin and ZO-1 expression, Nrf2 activation, and NQO1 and HO-1 expression.Zhang et al.  conducteCrocus sativus aqueous extract against this model of IBD. The animals presented elevated DAI, diminished body weight and colon length, augmented TNF-α and IFN-γ levels, and decreased HO-1, GPX-2, and Nrf2 expression and activity. After treatment, the mice presented elevated Nrf2 expression and activity, increased HO-1 and GPX-2, and decreased TNF-α and IFN-γ expression levels.Singh et al.  conductePrunus mahaleb concentrated fruit extract. Although the mice presented elevated intestinal inflammation and OS marked by decreased Nrf2 expression and activity, as well as diminished GSH, GSR, and glucose-6-phosphate dehydrogenase (G6PD) before treatment, the use of the fruit extract reverted this situation and augmented the Nrf2 expression and activity of the treated animals, as well as their expression of GSH, GSR, and G6PD.Ferramosca et al.  studied Quercus ilex L. leaf polyphenolic extract in the activation of the Nrf2 signaling pathway. Before treatment, the mice presented elevated anorexia, intestinal adhesions to adjacent organs, weight/length ratio of the colon, transmural necrosis, edema, diffuse inflammatory cells infiltration, ulceration, and crypt distortion, as well as diminished body weight. Additionally, the mice presented increased MPO, TNF-α, IL-1β, COX-2, iNOS, IκB-α degradation, p65 nuclear translocation, p50 nuclear translocation, NF-κB-mediated transcriptional protein activation, and MAPK  protein activation, in addition to decreased HO-1 and Nrf2-mediated transcriptional protein activation. After treatment with the plant and further increased Nrf2-mediated transcriptional protein activation and HO-1 overexpression, the mice presented decreased MPO, TNF-α, IL-1β, COX-2, and iNOS expression, IκB-α degradation, p65 and p50 nuclear translocation, NF-κB-mediated transcriptional protein activation, and MAPK  protein activation.Castejón et al.  studied Ziziphus spina-christi fruit extract in activating Nrf2 signaling. Before treatment, the mice presented elevated weight/length ratio of the colon, necrosis, ulceration, corrosion, hemorrhagic diarrhea, intestinal focal infiltration of inflammatory cells, crypt distortion, and decreased intestinal mucin concentration. The mice also presented elevated MPO, lipid peroxidation (LPO), NO, GSH, TNF-α, IL-1β, iNOS, COX-2, and NF-κB activation and signaling, in addition to decreased SOD, CAT, GSR, GPx, and Nrf2/HO-1 activation and signaling levels. After treatment with the fruit extract, there was diminished p38, MAPK, and VEGF-A expression, Bax and caspase-3-mediated apoptosis, MPO, LPO, NO, TNF-α, IL-1β, iNOS, and COX-2 expression, as well as increased GSH, SOD/, CAT, GSR, GPx and Nrf2/HO-1 activation and signaling. The treated mice also presented diminished NF-κB activation and signaling.Almeer et al.  studied Perilla frutescens extracts against this model of IBD. The animals presented diminished body weight and colon length, increased DAI, diarrhea, bloody stool, and intestinal infiltration of inflammatory cells. Before treatment, the animals also presented elevated COX-2, iNOS, cyclin D1, NF-κB activation and translocation, IκBα phosphorylation and degradation, p65 nuclear translocation, STAT3 signaling, TLR4, Interferon regulatory factor 3 (IRF3) (an inducer of interferon production), IFN-α, and IFN-γ, in addition to reduced levels of Nrf2/HO-1 activation and signaling and of HO-1. After treatment, the animals showed elevated levels of Nrf2/HO-1 activation and signaling and HO-1 expression, as well as decreased COX-2, iNOS, and cyclin D1 expressions, NF-κB activation and translocation, IκBα phosphorylation and degradation, p65 nuclear translocation, STAT3 signaling and TLR4, IRF3, IFN-α, and IFN-γ expression.Park et al., 2017  studied Zingiber officinale Roscoe, on colitis-associated Raw 264.7 macrophage cells, colon-26 LPS-activated cells, and a DSS-induced FVB/NJ mice model of colitis. Before treatment, the models presented elevated inflammation and OS measured by elevated levels of pro-inflammatory cytokines  and lipocalin (Lcn) 2. After treatment, the animals had activated Nrf2 signaling; therefore, the associated diseases were ameliorated by increased HO-1 levels and decreased TNF-α mRNA, IL-6 mRNA, IL-1β mRNA, and iNOS mRNA levels.Zhang et al., 2018  conducteGlycyrrhiza species, against Nrf2 deactivation during IBD. The mice presented intestinal morphological alterations due to increased inflammatory and oxidative damage. Furthermore, the rats showed elevated MPO, TNF-α, IL-1β, IL-6, COX-2, NF-κB p65, IKKα, p-IκB, NO, and Keap1 expression levels, as well as decreased SOD and GSH levels. After treatment, the mice presented activation of Nrf2 due to increased HO-1 and GCL and decreased Keap1, NF-κB- p65, IKKα, and p-IκB expression levels.Liu et al. , during Kim et al.  evaluateFerrari et al.  conducteLi et al.  conducteTan & Zheng  conducteWang et al.  evaluateJeon et al.  studied Pandurangan, Mohebali et al.  evaluateAlattar et al.  conducteChen et al.  conducteZhang et al.  studied Wu et al.  worked oYuan et al.  conducteQi et al.  studied Khodir et al.  considerKhater et al.  performeFibraureae caulis Pierre. During the study, after using DSS, there was an increase in DAI, weight loss, massive infiltration of inflammatory cells, destruction of crypts, thickening of the intestinal mucosa, and an increase in inflammatory mediators in the colon. There was an activation of NLRP3, an increase in TNF-a, IL-1B, IFN-γ, IL-17A, IL-6, MDA, MPO, ASC, caspase-1 and NO, and a reduction in IL-10 levels, TAC, SOD, GSH, CAT, GSH-Px, NRF2, and HO-1. The use of OPAL provided greater expression of mRNA and proteins of the Nrf2 and HO-1 pathways, in addition to inhibiting the activation of the NLPR3 inflammasome and the expression of mRNA of the NLRP3 proteins, ASC, and caspase-1. With the use of OPAL, there was also an attenuation of clinical manifestations, reduction in DAI, and less pathological damage and markers of OS and intestinal inflammation.Cheng et al. , during Wang et al.  performeRadix Ophiopogon japonicus widely used for treating inflammation and cardiovascular diseases, in the activation of Nrf2 to improve the function of the intestinal barrier. During the in vivo study with a DSS-induced C57BL/6J mice model of colitis, before the DSS administration, there was an increase in DAI, intestinal barrier dysfunction, abscesses, and crypt ulcers, higher levels of intestinal inflammation, apoptosis, and bacterial translocation and increased levels of TNF-a, IFN-y, NLRP3, caspase-1 and 11, GSDMD, Bax, and caspase-3. After using RUS, the mice had improved damage induced by DSS and more remarkable survival, in addition to activation of the Nrf2/NQO1 pathway, higher levels of B-cell lymphoma 2 (Bcl-2) protein, and attenuation of the Bax/c-caspase-3 pathway. During the in vitro study, the inflammatory injury was caused by LPSs in rat colon organoids. The organoids presented increased TNF-α and IFN-γ, factors related to macrophage migration, higher expression of Bax and caspase-3, increased apoptosis, and reduced Bcl-2 expression. On the other hand, with the use of RUS, there was an increase in the expression of Nrf2 and NQO1 proteins, as well as a reduction in the expression of TNF-α and IFN-γ and of factors related to macrophage migration.Using an in vivo and in vitro study, Wen et al.  evaluateWan et al. , during Schisandra chinensis. The in vivo study used a DSS-induced mice model of colitis which demonstrated increased DAI, formation of intestinal ulcers, dysfunction of intestinal integrity and permeability, greater bacterial translocation, inflammatory infiltration, and increased levels of TNF-α, IL-6, IL-18, and IL-1β before treatment. After using schisandrin B, there was a reduction in the inflammasome NLRP3 activation and in the production of TNF-α, IL-6, IL-18, IL-1β, caspase, and GSDMD with simultaneous activation of the AMPK/NRF2 pathway and reduction of ROS-induced mitochondrial damage. During the in vitro study, there was stimulation of inflammatory injury in human colon cancer cell line (HCT)-116 cells by LPSs, which caused an increase in the levels of TNF-α, IL-6, IL-18, IL-1β, NLRP3 activation, ROS, GSDMD, apoptosis, and mitochondrial damage, as well as increased lactate dehydrogenase (LDH) activity. With the use of schisandrin B, there was a reduction in all inflammatory cytokines expression that had previously been increased, in addition to greater expression of protein p-AMPK and Nrf2, with a reduction in oxidative mitochondrial damage and a relative improvement in colitis.W. Zhang et al. , during Garcinia kola nuts, against a mice model of colitis. The induction of colitis in rats with DSS caused weight loss, increased DAI, a reduced number of crypts, increased inflammatory infiltration, and increased synthesis of ROS and pro-inflammatory cytokines. After the administration of GB1a, there was a reduction in the NF-κBp65 translocation to the nucleus with inhibition of the NF-κB pathway, a decrease in the levels of MPO, MDA, IL-6, and TNF-α mRNA, and diminished levels of chemokines such as CCL5, CCL20, and CXCL1. Concomitantly, there was an increase in the translocation of Nrf2 to the nucleus and its expression, higher production of HO-1, GSH, and SOD, and higher levels of ZO-1 mRNA and occludin mRNA.Yu et al.  performeLi et al. , during Atractylodes macrocephala, including Atractylenolide III, in the TNBS-induced mice model of colitis. With TNBS, the animals presented increased DAI, intestinal inflammation, weight loss, colon shortening, infiltration of inflammatory cells, OS, submucosal necrosis, distortion of the mucosal structure, and ulcers, along with increased MPO, IL-1β, TNF-α, MDA, Formyl peptide receptor 1 (FPR1), NADPH (nicotinamide adenine dinucleotide phosphate hydrogen) oxidase 1 (NOX1), and phosphorylated Nrf2, as well as reduced levels of Nrf2 activation and signaling, SOD, GSH-Px, and CAT. After treatment with atractylenolide III, there was a reduction in the MPO, FPR1, NOX1, phosphorylated Nrf2, MDA and IL-1β, and TNF-α mRNA expressions, as well as increases in FPR1, Nrf2 activation, and signaling, CAT, SOD, and GSH-Px.Ren et al.  carried Polydatin, a bioactive compound isolated from the root and rhizome of Polygonum cuspidatum. Before treatment, the animals presented elevated DAI and OS, increased secretion of pro-inflammatory cytokines, and reduced colon length. After treatment, there was elevated nuclear translocation and phosphorylation of Nrf2, diminished phosphorylation of Erk1/2 and JNK1/2, and increased expression of HO-1, NQO1, and IL-10, as well as increased Akt phosphorylation. Peritore et al. [Polygonum cuspidatum but evaluated its roles against IBD in a DNBS-induced CD1 mice model of colitis. In this study, the mice presented, in addition to intestinal inflammation and OS, prominent transmural inflammatory cell infiltration and ulcer formation. Reduced NF-κB translocation, IkBα degradation, and elevated levels of SIRT1, Nrf2, and HO-1 expressions were highlighted with the treatment.G. Chen et al.  worked we et al.  also stuRosmarinic Acid against this model of IBD. The mice presented elevated DAI, intestinal inflammation, prominent transmural inflammatory cell infiltration, and elevated expression of inflammasome-related proteins. After treatment, the animals expressed elevated Nrf2 and HO-1 and reduced pro-inflammatory and OS genetic expression.Marinho et al.  conducteImperatorin, an active furocoumarin of Angelica dahurica, against this model of IBD. Before treatment, the animals presented elevated DAI, intestinal inflammation, and OS, coursing with increased TNF-α and IL-6 expression levels. The rats expressed elevated Nrf2 levels, increased ARE and HO-1, and reduced OS genetic expression with treatment.Luo & Luo  worked wJia et al.  conducteZingiberaceae and Araceae plants. Before treatment, the mice presented elevated intestinal inflammation and secretion of pro-inflammatory cytokines. After the use of curcumin, the authors reported an increase in the SIRT1/Nrf2 pathway, a reduction in TLR4 expression, and also a decrease in pro-inflammatory genetic expression.Yin et al.  worked wSesamin, there was a decrease in Keap1 levels, and it promoted an increase of the Nrf-2/ARE expression and induced the expression of p-AKT/AKT, GCLC, glutamate-cysteine ligase modifier subunit (GCLM), NQO1, and HO-1.Bai et al.  conducteMelia toosendan. This study used a DSS-induced C57BL/6 mice model of colitis. Before treatment, the mice presented elevated DAI, increased intestinal inflammation, and increased OS (elevated ROS). After using the compound, the animals presented decreased pro-inflammatory and OS gene expression as soon as toosendanin promoted an increase in Nrf2 and HO-1 expression levels.Fan et al.  worked wPropolis and Alpinia ofcinarum, known as galangin, against these models of IBD. The animals presented elevated DAI, the loss of epithelial and goblet cells, crypt aberrations, increased levels of pro-inflammatory cytokines (IL-6 and TNF-α), increased NF-κB activation, and elevated OS. After treatment, galangin induced an increase in the Nrf2 signaling pathway activation, as well as increases in HO-1, decreases in pro-inflammatory genetic expression, reductions in JNK phosphorylation, and diminished p-IKKα/β expression levels.Sangaraju et al.  studied Picrorhiza kurroa. The animals presented increased intestinal OS and intestinal inflammation, as well as elevated COX-2 expression. After using apocynin, the mice presented an increase in the Nrf2 signaling pathway activation, upper levels of HO-1, and decreased pro-inflammatory genetic expression.Hwang et al.  conducteGuo et al.  worked wRadix linderae. The animals presented elevated intestinal inflammation marked by increased levels of pro-inflammatory cytokines and an increased OS. After treatment, the rats expressed Nrf2 signaling pathway activation and decreased NLRP3 inflammasome activation.Lv et al.  conducteL. Yang et al.  worked wSeo et al.  studied Yang et al.  conducteCrespo et al.  worked wIn a mice model of IBD, Zhu et al.  evaluateIn a DSS-induced mice model of colitis, Zheng et al.  studied 2O2-induced OS and cell death via p38 MAPK-Nrf2/ARE pathway inhibition. These results suggested digitoflavone can be considered a protective factor against colorectal carcinogenesis.Yang et al.  studied Pandurangan, Saadatdoust, et al.  studied Tussilago farfara L., in experimental colitis-associated colon cancer. The authors used the DSS-induced BALB/c mice model of colitis. The administration of tussilagone diminished the prevalence of colonic tumors among the animals, reduced cancer cell proliferation, induced cancer cell apoptosis, and decreased β-catenin expression. This bioactive compound also exerted antioxidant effects against tumor formation due to increased HO-1 expression.Nam & Kim  studied Lippmann et al.  showed iKawabata et al.  studied Chiou et al.  intendedChiou et al.  evaluateWu et al.  studied The effects of wogonin were investigated by Yao et al.  in colorLong et al.  investigBagalagem et al.  exploredUlva pertusa, against Nrf2 deactivation in IBD. The treatment was carried out in the DNBS-induced CD1 mice model of colitis. Ulva pertusa showed immunomodulatory and anti-inflammatory properties by upregulating Nrf2 and inducing HO-1 activity. It also reduced COX-2 and iNOS and modulated the NF-κB pathway during the inflammatory burden of the animals’ intestines during IBD development. This seaweed also restored the damage caused by induced colitis, improved tissue restoration, reduced intestine edema, and reversed weight loss. Ardizzone et al.  studied Coriolus versicolor in a dose of 200 mg/kg against IBD. Before treatment, they observed neutrophils infiltration in the colon, lipid peroxidation, and increased pro-inflammatory cytokines secretion, such as IL-1β and TNF-α. Coriolus versicolor decreased the expression of TLR4 and reduced the NF-κB pathway activation and signaling, reverting the inflammatory and oxidative previous scenario. These actions were principally related to the Nrf2 activation and consequent HO-1 expression.Impellizzeri et al.  used a DSaccharomyces boulardii, a probiotic yeast resistant to low pH and highly tolerant to bile acids, in mice models of UC by regulating the NF-κB and Nrf2 signaling pathways. The authors used a DSS-induced C57BL/6 mice model of colitis and, after treatment with 105 and 107 CFU/mL, p.o. of S. boulardi, this probiotic reduced DAI effects and reestablished the gut health of the treated animals. Significantly, it also prevented IκBα degradation, suppressed nuclear translocation of NF-κB, reduced the activity of MPO, regulated inflammatory cytokines, and acted as an immune modulator. These actions were principally related to the Nrf2 signaling activation and the transcription of genes of antioxidant enzymes such as HO-1.Gao et al.  studied Gloeostereum incarnatum, a fungus rich in amino and fatty acids, on UC via modulation of Nrf2/NF-κB signaling in C57BL/6 mice. The use of the fungus reduced ROS and pro-inflammatory cytokines, such as IL-1β, IL-2, IL-6, IL-12, TNF-α, TNF-β, IFN-α, and IFN-γ. It also reduced the phosphorylated activation of NF-κB and regulated Nrf2/NF-κB signaling by principally decreasing CAT, HO-1, SOD-1, and SOD-2.Li et al.  exploredBee pollen has polyphenolic compounds and provides various health benefits, mainly by its anti-inflammatory properties. Li et al.  studied Jenkhetkan et al.  exploredNumerous plants and phytochemicals can be used as therapeutic adjuvants in IBD and colorectal cancer associated with IBD since they can regulate Nrf2 and, therefore, among many actions, reduce inflammation and OS and increase apoptosis of cancer cells. These effects improve the bowel environment, mucosal barrier, colon, and crypt disruption, reduce ulceration and microbial translocation, and consequently reduce the disease activity index. Moreover, the modulation of Nrf2 can regulate various genes involved in cellular redox, protein degradation, DNA repair, xenobiotic metabolism, and apoptosis, contributing to the prevention of colorectal cancer.Even though only in vivo and in vitro studies were found in the consulted databases, our results bring to light the need for clinical trials so that the effects on human beings can be investigated and new paths can be paved in the search for effective therapy (or effective adjuvant therapy) for IBDs, whose incidence increases rapidly and drastically reduces the quality of life and capacity for work and social interaction of the carriers.As a limitation, our study is not different from other narrative reviews. Due to the significant number of included studies, determining and integrating complex interactions between different models of IBD and IBD-associated colorectal cancer presented to be a challenge. However, this could be overcome by the objective of this review, which was to respond to the question, “What are the effects of medicinal plants and natural and synthesized phytochemicals as regulators of the Nrf2 pathways in IBD and colitis-associated colorectal cancer?’’ and giving to light the need for clinical trials.The present review is the first in the medical literature to evaluate the roles of medicinal plants and natural and synthesized phytochemicals in Nrf2 regulation during IBD and IBD-related colorectal cancer. In addition, to our knowledge, Nrf2 regulation in IBD and IBD-associated carcinogenesis was never before revised in light of a narrative review that organized the databases’ model studies in tables with our included topics.


We reviewed the literature on the toxic effects of AFB1 in humans and animals along with its toxicokinetic properties. The damage induced by AFB1 in cells and tissues is mainly achieved through cell cycle arrest and inhibition of cell proliferation, and the induction of apoptosis, oxidative stress, endoplasmic reticulum (ER) stress and autophagy. In addition, numerous coding genes and non-coding RNAs have been identified that regulate AFB1 toxicity. This review is a summary of the current research on the complexity of AFB1 toxicity, and provides insights into the molecular mechanisms as well as the phenotypic characteristics.Fungal contamination of animal feed can severely affect the health of farm animals, and result in considerable economic losses. Certain filamentous fungi or molds produce toxic secondary metabolites known as mycotoxins, of which aflatoxins (AFTs) are considered the most critical dietary risk factor for both humans and animals. AFTs are ubiquitous in the environment, soil, and food crops, and aflatoxin B Aspergillus flavus and A. parasiticus, and is a derivative of difuranoxano-naphtho-ketone consisting of a difuran ring and a coumarin ring. AFTs are produced at temperatures between 10–40 °C, pH 3.0–8.5, moisture content 18–20%, and water activity > 0.82 , and AFG1 and AFG2 (green) types , AFP1, AFQ1 and AFH1 are derivatives of AFTs. AFB1 is considered to be the most toxic AFT  types . AFM1, Aoxic AFT , with 10oxic AFT , and is oxic AFT . AFB1 hafunction , decreasfunction , and resand milk . Long-teand milk . Death dountries . Studieslbladder , and 4.6lbladder .1 poisoning, there is no systematic review on the hepatotoxicity, enterotoxicity, nephrotoxicity, immunotoxicity, neurotoxicity and reproductive toxicity of AFB1. In this review, we have discussed the mechanisms under the toxic effects of AFB1 in order to provide a reliable reference for further research in animal husbandry, mycotoxins, and the treatment of toxin-related diseases in humans, livestock and poultry.Despite reports of liver toxicity and immunotoxicity in humans and animals due to AFBPubMed and Web of Science databases were searched for relevant articles. A total of 7,715 articles appeared Pubmed database using “aflatoxin B1” as the search term and the date of publication from 1975/1/1 to 2022/3/1 . After narrowing the search with keywords including “toxicokinetics of aflatoxin B1”, “hepatotoxicity of aflatoxin B1”, “enterotoxicity of aflatoxin B1”, nephrotoxicity of aflatoxin B1, “neurotoxicity of aflatoxin B1”, “immunotoxicity of aflatoxin B1”, “reproductive toxicity of aflatoxin B1”, “aflatoxin B1 and COX2”, “aflatoxin B1 and Nrf2”, “aflatoxin B1 and p53”, “aflatoxin B1 and microRNA”, “aflatoxin B1 and lncRNA”, and “aflatoxin B1 and livestock”, 1,733 studies were obtained. With “aflatoxin B1” as the search term, 11,741 articles published between 1975–2022 appeared in the Web of Science database, of which 1,514 articles were selected as above. After removing duplicate articles and the articles with little relevance, 137 articles were selected for the review.1 exposure occurs through dietary intake, skin contact and inhalation of contaminated dust. More than 80% of the ingested AFB1 is absorbed in the duodenum and the jejunum through passive transport , which includes the endo-8,9-epoxide (endo-AFBO) and exo-8,9-epoxide (exo-AFBO) isomers  , such as hydroxyl free radicals, hydrogen peroxide and other free radicals that damage cell membrane and macromolecules   that is B1-FAPy) . However protein . In addiodon 248 . The p53odon 248 . Furtherolecules . The com1 differs across species. Monogastric animals are more susceptible to AFB1 compared to the ruminants since the gut microbes in the latter can metabolize mycotoxins. Long-term exposure to low levels of AFB1 is more common in livestock as opposed to acute poisoning , aspartate aminotransferase (AST), alkaline phosphatase (ALP), serum creatinine (SCR), catalase (CAT) and malondialdehyde (MDA) in the affected cattle, and decreased that of total protein (TP), magnesium (Mg) and glutathione (GSH). Furthermore, autopsy of the poisoned cattle showed hepatomegaly, gallbladder enlargement, and intestinal and renal hyperemia  in the liver cells, leading to oxidative stress, inflammation, and liver damage   signaling pathway  . Prolongtabolism , and restabolism . Likewishe liver . In one  pathway . Epidemir cancer . Furtherr cancer . To summ1 has been associated with chronic intestinal diseases. AFB1 caused intestinal mucosal injury and inhibited IECs proliferation in mice  patients. The combination of metabolic dysfunction, loss of IECs and glandular atrophy caused by AFB1 can lead to chronic intestinal diseases  . Long-te in mice . In addiractions . AFB1 albacteria . The micdiseases .1. Broilers fed with different doses of AFB1 exhibit severe damage to the intestinal villi characterized by lower density and absorption area  molecules and apoptotic factors  and average daily feed intake (ADFI), resulting in reduced growth and development. Furthermore, the relative weight of the digestive organs, the activity of digestive enzymes and the biochemical indices of intestinal development were also altered   . AFB1 exelopment . In addipoptosis . To summ1-induced immunotoxicity  in the liver cells  and human monocyte-derived dendritic cells (MDDCs), and increased apoptosis  and decreasing GSH levels. These changes correlated with upregulation of NOS2, TNFα and CXCL2 mRNAs, and downregulation of CD86. AFB1-induced oxidative stress in macrophages also impaired the mitochondrial respiratory chain, leading to activation of the inflammatory response pathways  increased production of TNFα and IL6 in these cells, and decreased lactate dehydrogenase secretion and the phagocytotic index in a concentration-dependent manner. In addition, the combination treatment significantly decreased the production of GSH, increased ROS levels, and promoted IκBa degradation, NF-κB phosphorylation and nuclear translocation of NF-κB. Thus, AFB1 and OTA can synergistically aggravate immunotoxicity by activating of the NF-κB signaling pathway , and consumption of AFB1-contaminated feed increased the risk of secondary infections in pigs (1 activated the release of heterophil extracellular traps (HETs) in chicks, and induced the expression of TNFα, IL6 and IL1β, iNOS, COX2, NLRP3, caspase1, caspase3 and caspase11, which resulted in liver and kidney damage  have been identified in the liver tissues of AFB1-treated versus healthy mice. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the DEGs were enriched in functions such as cell adhesion, cell proliferation and cell cycle regulation   . Finallypathways .1 can alter the expression levels of oncogenes (such as ras and c-fos) and tumor suppressor genes (such as p53 and survivin) (1 induced HCC cases (1 (AFBurvivin) , and indCC cases . In a recases  is a key mediator of AFBtoxicity .1 induces COX2 expression, promotes mitochondrial autophagy and impairs mitochondrial lipid metabolism in hepatocytes, leading to hepatic steatosis  and ER stress-response genes  in macrophages  , gga-miR-190a-3p and Bcl-6 gene induce apoptosis in liver cells, whereas co-expression of lncRNA (TU10057), gga-miR-301a-3p, gga-miR-301b-3p and PPARG gene cause fatty liver  . MiRNA-4 pathway . AFB1-inty liver . Taken togenesis .1 is a major health concern. In this review, we summarized the hepatotoxic, enterotoxic, nephrotoxic, neurotoxic, immune-toxic and gonado-toxic effects of AFB1. In addition, the mechanisms underlying the toxicity of AFB1, including cell cycle arrest, apoptosis, oxidative stress, ER stress and autophagy, and the effector genes, non-coding RNAs and signaling pathways were also discussed.The contaminated of agricultural produce with AFTs is practically unavoidable worldwide. The presence of AFTs in feeds may decrease feed intake, damage health and affect livestock productivity. In addition, the toxic residues in animal products  may have some adverse effects on human health. Contamination of plant and animal-derived food products by AFB1 is very complex, and is closely related to the dose, exposure duration, administration mode, solvent, species, gender, age, target organs and so on. More comprehensive and systematic tests are needed to ascertain the acute and chronic AFB1 exposures doses in different species, develop standard dosing methods, and determine the quantified dose–response relationship in vitro and in vivo. In addition, the combined effects of AFB1 with other mycotoxins and with macronutrient deficiency, and well as the underlying mechanisms will be our future research concerns. The relationship between gut microorganisms and sensitivity to AFB1, and the impact of intestinal microbiota on AFB1 metabolism will also be an important research focus. Elucidating the molecular mechanisms underlying the toxicity of AFB1 can help mitigate or eliminate the toxic effects of AFB1 by genetic methods, which will also be worth investigating.The toxicity of AFB10.7717/peerj.13850/supp-1Supplemental Information 1The number list of articles searched via databases.Click here for additional data file.


Developmental and epileptic encephalopathies (DEEs) have high genetic heterogeneity, and DEE due to the potassium voltage-gated channel subfamily C member 2 (KCNC2) variant remains poorly understood, given the scarcity of related case studies. We report on two unrelated Chinese patients, an 11-year-old boy and a 5-year-old girl, diagnosed with global developmental delay (GDD), intellectual disability (ID), and focal impaired awareness seizure characterized by generalized spike and wave complexes on electroencephalogram (EEG) in the absence of significant brain lesions. Whole-exome sequencing (WES) and electrophysiological analysis were performed to detect genetic variants and evaluate functional changes of the mutant KCNC2, respectively. Importantly, we identified a novel gain-of-function KCNC2 variant, R405G, in both patients. Previously reported variants, V471L, R351K, T437A, and T437N, and novel R405G were found in multiple unrelated patients with DEE, showing consistent genotype–phenotype associations. These findings emphasize that the KCNC2 gene is causative for DEE and facilitates treatment and prognosis in patients with DEE due to KCNC2 mutations. KCNC2) gene and DEE remains poorly understood due to the scarcity of reported cases  is a group of epilepsy syndromes characterized by severe seizures that are drug-resistant and significant developmental delay or regression . Most paed cases .KCNC2 gene, also known as KV3.2, together with KCNC1, KCNC3, and KCNC4 (KV3.4), belongs to the Shaw-related potassium channel family (KV3 subfamily). It is well-established that KV3 proteins share the structure of six-transmembrane (6-TM) voltage-gated K + channels, which maintain the potassium ion permeability of excitable membranes by regulating pore penetration in 6-TM in response to voltage changes, thereby maintaining neuronal functions, such as maintenance of membrane potential, regulation of cell volume or neurotransmitter release, and modulation of electrical excitability  by regulating the membrane resting potential and fast action  in mice resulted in a deficit of high-frequency oscillations modulation, distorted cortical rhythmic activity, suppressed cortical inhibition, impaired fast spiking in cortical interneurons, enhanced susceptibility to epileptic seizures and abnormal sleep patterns, and increased anxiety in the open field. However, it can be challenging to discern whether a neurodevelopmental delay has occurred , in two unrelated patients who were both clinically diagnosed with DEE. We demonstrated that the variant R405G yielded higher activity in vitro than the wild-type (WT) KCNC2 on a range of functioning voltages.In this study, we identified a novel heterozygous This study was approved by the Ethics Committee of Wuhan Children’s Hospital, Hubei, China . Written informed consent for genetic testing and publication of photographs was obtained from the legal guardians of each subject. All investigations were conducted in accordance with the principles of the Declaration of Helsinki.Two patients from unrelated families were admitted to the Department of Neurology, Wuhan Children’s Hospital. The medical records, including medical history, symptoms and signs, physical examinations, laboratory tests, radiological results, and developmental assessment, were documented by the attending physicians. Genetic tests and analyses were performed by Chigene .2 including allele frequency in the Single Nucleotide Polymorphism database (dbSNP), 1000 Genomes Project, Exome Aggregation Consortium (ExAC), Genome Aggregation Database (gnomAD), ESP, and Chigene in-house MAFs database; positions and associated clinical phenotypes based on UCSC, RefGene, GENCODE, and ENSEMBL transcripts; LOVD, SWISS, Clinvitae, HGMD, OMIM, and ClinVar databases; and functional and conservational predictions for the amino-acid changes according to software packages . Variant filtering was done according to rare variants with minor allele frequency (MAF) of <1% and biological impact, gene features and disease mechanism, recessive/dominant/de novo pattern of inheritance and segregation analysis, phenotypic relevance through human phenotype ontology (HPO) terms, and scientific literature. Subsequently, pathogenic or likely pathogenic variants were evaluated according to the American College of Medical Genetics (ACMG) practice guidelines.Whole-exome sequencing (WES) was performed on the proband’s genomic DNA extracted from peripheral blood leukocytes using xGen Exome Research Panel version 1.0  and paired-end sequencing (2 × 150 bp) on NovaSeq 6000  with not less than 99% regions covered at least 10×. Raw sequence readouts were processed by fastp. Paired-end sequence reads were aligned to the human GRCh37/hg19 genome with Burrows–Wheeler Aligner (BWA). Initial mapping process and variant site identification were conducted using GATK. Annotations were retrieved from the database-based online system independently developed by Chigene ,The KCNC2 (NM_139137) variant c.1213A > G treated as plausible disease-causing was verified in the probands and their parents by Sanger sequencing performed using the BigDye Terminator version 1.1 Cycle Sequencing Kit on an ABI 3730XL instrument  to define the inheritance model. The primers used are 5′-CTGACCATGTTTGGGGGTACA-3′ and 5′-GAAACGGATCCTGCCTTGAC-3′ . PCR amplification was carried out with a KAPA2G Robust HotStart PCR Kit  on a Hema 9600 PCR Thermo Cycler . The DNA sequences were analyzed using the DNASTAR software package.Predicted (AF-Q96PR1-F1) configurations of human KCNC2 in the AlphaFold Protein Structure Database served as the template for developing a structural model that covered all 638 residues (NP_631875.1). Hydrogen-bond networks of the WT and the variant proteins were visualized in PyMOL.2 atmosphere. HEK-293 cells were seeded in six-well plates and subsequently transfected with 2 μg of WT or mutant cDNAs by transient transfection using Lipofectamine 3000 . The day after transfection, cells were seeded into a 24-well plate at a density of 8 × 103 cells per well. Approximately 48 h after transfection, cells were selected based on the intensity of the fluorescent reporter and placed in the recording chamber.HEK293 cells were cultured at 37°C in DMEM supplemented with 10% fetal bovine serum (FBS)  in a humidified 5% (v/v) CO2.6H2O, 2 CaCl2.2H2O, 10 D-glucose, 10 N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES), 1.25 NaH2PO4.2H2O, and pH adjusted to 7.4 with sodium hydroxide (NaOH). The pipette solution contained (in mM) 20 KCl, 115 K-aspartic, 1 MgCl2.6H2O, 5 ethylene glycol tetraacetic acid (EGTA), 10 HEPES, 2 Na2-ATP, and pH adjusted to 7.2 using potassium hydroxide (KOH). After the break-in, a 5-min waiting period equilibrated the cytoplasm of the cell and the pipette solution. Currents were activated by 500 ms voltage pulses ranging from −80 to +80 mV (in 10 mV increments) from a holding potential of −80 mV at a sweep frequency of 100 ms, calculated by dividing the current (in pA) by the cell capacitance (in pF). Current–voltage curves were converted through a Boltzmann equation (g(V) = gmax/(1 + exp((V-V0.5)/k))), where g, conductance; gmax, maximal conductance; V, voltage; V0.5, voltage of half-maximal activation; and k, slope factor into normalized conductance–voltage curve.The macroscopic current of transiently transfected HEK-293 cells was recorded with an EPC10 amplifier  and the Patchmaster software using the whole-cell configuration of the patch-clamp technique. The sampling interval was set at 100 ms (20 kHz) in all experiments. Pipettes  were pulled from borosilicate glass and had resistances between 1.5 and 2.5 MΩ when filled with the pipette solution. The glass pipette was pulled using a micropipette puller and manipulated using a micro-manipulator under the microscope. After touching the cell, a slight suction was applied to achieve high seal resistance (GΩ). Fast capacitance (in pF) compensation was made after achieving a high seal, and the membrane was broken. Cell capacitance (in pF) compensation was made from whole-cell capacitance compensation after the whole-cell mode was achieved. No leak subtraction was made. The extracellular solution contained (in mM) 140 sodium chloride (NaCl), 3.5 potassium chloride (KCl), 1 MgClt-test comparison. Significant differences were assessed by Student’s t-test, with *p < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.Data are expressed as mean ± standard error of the mean (SEM). At each measured voltage, e.g., −40, +10, and +60 mV, the membrane currents of mutant and WT cells were repeatedly tested three times, taking the mean value and SEM for V3.2,” “developmental and epileptic encephalopathy,” “DEE,” or “epilepsy” were used for literature search in the PubMed, Medline, and Google Scholar databases.Keywords “KCNC2” or “KPatient 1 was a 9-year-old boy admitted to our hospital for intractable epilepsy. He was born to a non-consanguineous Chinese couple with a history of birth asphyxia and hypoxia. The parents claimed no family history of seizures. At the age of 3 months, the patient experienced paroxysmal attacks with rolling back of eyeballs and loss of consciousness several times per day, which led to an initial diagnosis of focal epilepsy at the local hospital. For the following 8 years, he was successively treated with levetiracetam (LEV), valproate acid (VPA), oxcarbazepine (OXC), lamotrigine (LTG), and nitrazepam (NZP), which were ineffective for seizure control. Despite normal growth, the patient had intellectual disability (ID) with language and motor impairment. At the age of 9 years, the patient could not run and jump and had moderate ID (IQ 56). He experienced several focal seizures with episodic atypical absences during his hospitalization. Electroencephalogram (EEG) showed slowing of the basic background rhythm with hypsarrhythmia and generalized multifocal polyspike and slow wave, especially in the sleep stage. During a seizure, EEG showed generalized high amplitude waves at 3–4 Hz and spike-and-slow-wave bursts for 8–18 s. His blood and urine tests and brain MRI were normal. The patient was seizure-free after treatment with a combination of VPA/LTG/NZP. At the last follow-up, the patient (at the age of 11 years) was seizure-free with multifocal slow-wave, sharp-slow-wave, sharp-wave, and spike complexes on awake EEG. Complexes of multifocal polyspike and slow wave, spike and slow-wave, and sharp-wave were conducted by forward modeling EEG during sleep .Patient 2 was a girl aged 4 years and 9 months and the first child of a non-consanguineous normal Chinese couple. The patient experienced severely delayed neuromotor development , with no ability to speak, and had severe ID (IQ 34). When she was 1 year and 6 months old, the parents found she was subject to spontaneous attacks of eyelid myoclonia  more than 10 times per day and myoclonic seizures (jerking hands), which usually lasted for 2–3 min and often occurred before sleeping. After the patient was diagnosed with focal onset epilepsy at the local hospital based on an EEG (images were unavailable). on EEG at the local hospital, an antiepileptic treatment was suggested to her. After a poor response to OXC, her seizures were relieved with a combination of LEV/VPA and she was seizure-free last year. At the age of 3 years, the patient came to our hospital, and she was diagnosed with global developmental delay (GDD) with focal epilepsy. EEG showed slowing of the basic occipital rhythm, multifocal generalized complexes of spikes, spike and slow-wave, polyspike and slow-wave, and sharp-wave, significantly at rest and during sleep. Her brain MRI was normal. When the patient was 4 years and 9 months old, upon the last follow-up, the EEG showed the same pattern of slowing of the occipital basic rhythm, multifocal generalized complexes of slow-wave, spike and slow-wave, sharp-slow-wave, and polyspike and slow-wave, especially in occipital, frontal pole, and frontal regions .KCNC2 variants had been documented in several reported DEE cases (de novo (Whole-exome sequencing was performed on both patients. No pathogenic or likely pathogenic variants were found in well-known DEE-associated genes from the OMIM database (OMIM phenotypic series: PS308350). However, we found a heterozygous KCNC2 variant in both patients, NM_139137.4: c.1213A > G (p.R405G). A review of the literature showed that EE cases , while R, KCNC2 contains cytosolic N- and C-termini and six membrane-spanning segments. KCNC2 R405 is located within the S5-S6 linker, an extracellular loop that forms one of the ion-selective macromolecular protein pores in KCNC2 . In addind KCNC4 . Analysind KCNC4 . As a reV3.2 channel functioning, we constructed EGFP-fusion expression vectors expressing KCNC2 WT and R405G cDNA sequences transfected into HEK-293 cells. Compared to WT, R405G caused a significantly higher density current at voltages between −40 and +10 mV  (To investigate the impact of R405G in Kd +10 mV . At +60 d +10 mV . By meas= 21.36) .de novo, except S333T in patient 13  to a certain extent, than patients with significant findings on brain imaging due to KCNC2-DEE . Among them, synaptopathies and neurological channelopathies are two large groups. DEEs caused by synaptopathies gene variants are usually highly heterogeneous and lack clear genotype–phenotype associations, but neurodevelopmental disorders characterized by central vision impairment due to VAMP2 deficiency constitutes the SNARE complex and hand motor loss in Rett-like syndrome, and myoclonic seizures and subsequent ataxia caused by STX1B pathogenic variants are beneficial to clinical differential diagnosis ; neuroloCACNA1H . EpileptEE cases , suggestKCNC2 variants on four sites, R405, T437, R351, and V471, which found amino acid changes in multiple unrelated individuals. The results revealed a remarkable genotype–phenotype association in these cases. In our study, two unrelated patients shared phenotypes, including focal impaired awareness seizure, motor developmental disorder, severe ID, and good response to antiepileptic drugs; in the case of V471, both patients had infantile spasms, severe ID, and ineffective antiepileptic medication  and loop , in which three interactions  could not be detected in the G405 mutated protein. Thus, the R405G variant would destabilize the channel by impeding such electrostatic interactions. Recently, the changes described for the R405G channel may differ from D167Y and V471L variants associated with DEE. D167Y is close to the N-terminal region (BTB/POZ domain) and may impair the tetramerization of the channel , while Vin vitro electrophysiological analysis, and the results revealed that KCNC2 R405G decreased maximal current density but shifted membrane potential to more negative values in the activation curve of KV3.2 channels. Currents carried out by R405G mutant were inhibited, exhibiting reduced selectivity for K+ ions, consistent with the location of the R405 residue in the pore helix before the selectivity filter, which interacted with residues in both S5 and pore helices to modulate the selectivity of KV3.2 channels. During the analysis of the channel kinetics, the R405G mutant displayed a significantly slower deactivation and significantly reduced current compared to WT KCNC2 channels (gain-of-function), which exhibit the same electrophysiological characterization as the D167Y mutant .The original contributions presented in this study are publicly available. Information about KCNC2 variant R405G reported in this study can be accessed online using id #0000872152 through the lovd database . Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin. Written informed consent was obtained from the individual(s), and minor(s)’ legal guardian/next of kin, for the publication of any potentially identifiable images or data included in this article.SW: designed and conceptualized the study, contributed to the revision of the manuscript, and editing. YY: designed and conceptualized the study. XW: recruited and phenotyped the patients. XD: contributed to the revision of the manuscript. ZL: drafted the manuscript and prepared the figures. JM: conducted electrophysiological studies. WG: conducted molecular analysis and interpreted the data. DS: conceptualization, visualization, resources, supervision, writing – review & editing, and funding acquisition. All authors reviewed the manuscript and agreed to the published version of the manuscript.