Parthenium hysterophorus Linn phytochemically and pharmacologically. Phytochemical screening is important for the isolation of active compounds before bulk extraction. The crude extracts and their fractions were screened for enzyme  inhibition assays, in vivo analgesic, anti-inflammatory, and sedative effects. Results indicated the presence of steroids, flavonoids, etc. The crude extracts such as methanol, hexane, aqueous, ethyl acetate, chloroform, and butanol exhibited excellent urease inhibitory activities with IC50 = 43.1 ± 1.24, 31.9 ± 2.21, 31.9 ± 2.21, 57.3 ± 1.27, 49.2 ± 2.16, and 35.3 ± 1.12, respectively, as compared to standard acetohydroxamic acid (20.3 ± 0.43). The extracts  also showed promising α-glycosidase potency with IC50 = 13.1 ± 0.34, 21.2 ± 1.16, 23.1 ± 0.12, 84.2 ± 2.17, 118.6 ± 3.07, and 840 ± 1.73, respectively against acarbose (840 ± 1.73). The phosphodiesterase activity of the mentioned extracts was also excellent with IC50 = 131.1 ± 2.41, 197.2 ± 3.16, 24.2 ± 0.11, 62.4 ± 2.21, 152.4 ± 1.81, and 55.3 ± 2.15, respectively, against the standard (265.5 ± 2.25). Furthermore, butanol (14.96 ± 1.78), ethyl acetate (18.98 ± 1.71), and methanol (16.87 ± 1.00) showed dose-dependent analgesic effects with a maximum inhibition of acetic acid-induced writhes. Whereas, methanolic and butanol extracts exhibited maximum inhibition of inflammation in the carrageenan paw edema test. The aqueous (p < 0.01) and butanol (p < 0.01) extracts exhibited maximum a sedative effect followed by chloroform (p < 0.05), ethyl acetate (p < 0.05), and methanolic (p < 0.05) fractions as compared to the standard drug. The current research concluded that Parthenium hysterophorus Linn has important phytochemical constituents having inhibitory effects on urease, α-glycosidase, and phosphodiesterase enzymes. Furthermore, the plant has analgesic, anti-inflammatory, and sedative effects. The P. hysterophorus needs to further be explored for the candidate molecules responsible for the abovementioned activities.The main aim of this research was to explore  Parthenium hysterophorus L.  is a flowering plant and an aggressive ubiquitous annual herbaceous weed belonging to the family Asteraceae. P. hysterophorus is commonly known as altamisa, bitter weed, white top, and carrot grass. The flowering of this plant occurs throughout the year [P. hysterophorus thrives in Pakistan, America, Africa, Asia, as well as in Australia. It has multi medicinal traditional usage such as for the treatment of fever, neurologic disorders, diarrhea, dysentery, malaria, urinary tract infections, allergic respiratory problems, mutagenicity in humans, and emmenagogue. In addition, P. hysterophorus has been employed in traditional medicine as a remedy for inflammation and rheumatism, and as an analgesic in muscular rheumatism [P. hysterophorus is not well-explored for its phytochemical constituents. Therefore, research is required for the isolation, purification, and structure determination of active constituents of this plant. P. hysterophorus has been reported to contain toxins called sesquiterpene lactones such as the glycoside parthenin [p-coumaric, p-hydroxybenzoin, vanillic acid, chlorogenic acid, anisic acid, ferulic acid, and various alcohols [P. hysterophorus have led to the isolation of fourteen compounds and some of them having cytotoxic potential [P. hysterophorus [P. hysterophorus, and to extract and isolate more active phytochemicals that rationalize its properties and pharmaceutical applications. The present study is an attempt to evaluate the in vitro enzyme  inhibition assays, in vivo analgesic, anti-inflammatory, and sedative effects of the various fractions of P. hysterophorus.the year , 2. P. heumatism . This plarthenin . Other palcohols . Ether aotential . A novelrophorus . Anotherrophorus –10. On tP. hysterophorus was collected from various areas of the University of Swabi, Anbar, K.P., Pakistan. The plant specimen was validated by a botanist in the Department of Botany, University of Swabi KP, Pakistan, and voucher specimen NO. BOT.UOS4 was deposited in the said department. The plant material was dried under shade at room temperature and on the ground to obtain a powder for extraction.Plant material of P. hysterophorus powder material was subjected to cold extraction using a polar organic solvent such as methanol or ethanol, distilled water, and n-hexane. The extract was concentrated by means of a rotary evaporator at a low temperature (50–55°C). This was followed by fractionation using organic solvents of different polarities such as Hexane, CHCl3, EtOAc, and methanol (×3 for each). The extract was then suspended with the minimum amount of water, followed by the addition of different organic solvents starting from nonpolar (n-hexane) to more polar ones such as chloroform, dichloromethane, ethyl acetate, and butanol, respectively. Each collected fraction was concentrated under a vacuum by using a rotary evaporator at a low temperature (50–55°C) to obtain a crude fraction. The crude extracts and fractions were subjected to phytochemical screening before bulk extraction. Finally, the obtained crude extracts and fractions were subjected to in vitro and in vivo biological assays.The obtained A phytochemical screening test of extracts/fractions was carried out for identification of active phytochemicals as per reported procedures –15.µL of crude extracts and their fractions (0.5 mM) and 25 µL urease catalyst (1 U/well) were hatched for 15 minutes at 30°C. Then, 55 µL substrate urea (100 mM) was re-brooded at 30°C for 15 minutes. After completion of incubation, 70 µL of alkali reagents (0.1% sodium hypochlorite) and 45 µL of phenol  were mixed. The incubation of the plates was again performed for 50 minutes at 30°C. The urease screening test was periodically done with continuous urea hydrolysis and ammonia production. The change in absorbance ) was monitored at 630 nm on an ELISA plate reader .The urease inhibition activity of the crude extracts and fractions was performed by spectrophotometry in 96-well plates as per the standard method . 5 µL of3)2CO (acetone) powder in normal saline  was sonicated appropriately, and the supernatant was used as a source of basic intestinal α-glucosidase after centrifugation [α-glucosidase for 5 min before 50 mL of substrate (5 mM p-nitrophenyl-a-D-glucopyranoside (p-NPG) was arranged in the similar buffer) was included. The α-glucosidase-mediated conversion of p-NPG into D-glucose and p-nitrophenol at 405 nm was monitored spectrophotometrically every 5 minutes. Singular seats for the screening extract were set up to extract baseline absorbance of the substrate and altered with 50 mL of buffer. The control sample contained 10 mL DMSO alongside screening samples. The percentage of enzyme inhibition was assessed as follows:A speaks to the absorbance of the control exclusive of the prepared extract sample, and B connects to absorbance in the attendance of test samples.The rat intestinal  and then used in the following calculation:This enzyme assay was performed using snake venom-derived PDE-1 (Sigma P-4631), adopting already published methods with some modifications . 30 mM Mn = 6) weighing 18–22 g were used. All animals were withdrawn from food 3 hours before the start of the experiment and the mice were distributed in different groups. Among the divided animals, group I was injected with normal saline (10 ml/kg) as the control, while group II was administrated with the standard drug , and the rest of the groups were administered with various extracts and fractions . After administration of 30 min, the animals were treated with 1% acetic acid. Then, the number of abdominal constrictions (writhes) was counted after 5 min of acetic acid injection for the period of 10 minutes as per the usual methods [BALB/c mice of both sexes  were treated with normal saline (10 ml/kg), standard drug, and tested extracts and their fractions (5 and 10 mg/kg i.p). The animals were introduced at one edge of the tube and then permitted to move up to the mark 20 cm from the base. When the treated animals touched the 20 cm mark, the tube was moved straight to the perpendicular position and the animals strained to climb again to the tube with a backward effort. The mouse which failed to reach up to the mark within 30 seconds was considered to have relaxed muscles [The crude extracts and various fractions of Parthenium hysterophorus is given in α-glucosidase, and phosphodiesterase inhibitory effects.Phytochemical analysis of Parthenium hysterophorus showed excellent urease inhibition activity. The polar extracts such as ethyl acetate (87.3%), butanol (84%), and aqueous (81.4%) showed excellent activity with IC50 values of 57.3 ± 1.27, 35.3 ± 1.12, and 31.9 ± 2.21, respectively. The inhibitory potential is followed by n-hexane (77.4%), methanolic (74.2%), and chloroform (74.2%) fractions which showed moderate activity with IC50 values of 39.8 ± 0.36, 43.1 ± 1.24, and 49.2 ± 2.16, respectively. Acetohydroxamic acid (96.3%) is used as a standard urease inhibitor with an IC50 value of 20.3 ± 0.43.The urease inhibitory activities of this plant are given in α-glycosidase inhibitory activities of this plant are given in P. hysterophorus showed excellent α-glycosidase inhibition potential. The extracts such as methanolic (94.2%), aqueous (91.4%), and butanol (90.2%) showed excellent activity with IC50 values of 13.1 ± 0.34, 23.1 ± 0.12, and 840 ± 1.73, respectively. The inhibitory potential of α-glucosidase was followed by n-hexane (84.7%), ethyl acetate (82.7%), and chloroform (76.4%) fractions with IC50 values of 840 ± 1.73, 84.2 ± 2.17, and 118.6 ± 3.07, respectively. The standard α-glucosidase inhibitor, acarbose, exhibited percent inhibition (90.2%) with an IC50 value of 840 ± 1.73.The P. hysterophorus showed excellent phosphodiesterase inhibition potential. The extract such as aqueous (91.4%), butanol (88.4%) and n-hexane (82.7%) showed excellent activity with IC50 values of 24.2 ± 0.11, 55.3 ± 2.15, 197.2 ± 3.16, respectively. The inhibitory potential of phosphodiesterase is followed by the methanolic (79%), chloroform (78.4%), and ethyl acetate (76.5%) with IC50 values of 131.1 ± 2.41, 152.4 ± 1.81, and 62.4 ± 2.21, respectively. The standard phosphodiesterase inhibitor, EDTA exhibited 87.9% inhibition with an IC50 value of 265.5 ± 2.25.The phosphodiesterase inhibitory activities of this plant are given in P. hysterophorus including methanol, hexane, aqueous, ethyl acetate, chloroform, and butanol were assessed for analgesic activity. All the extracts/fractions were tested at 25, 50, and 100 mg/kg and the effects were observed to be dose-dependent. Among the tested extracts, butanol, ethyl acetate, and methanol showed maximum analgesic effect as compared to the standard drug , flavonoids, and various alcohols [P. hysterophorus [Phytochemicals in medical plants play key roles in their biological potency. Phytochemical analysis plays a significant role in the isolation of new active and rare compounds. The biomedical importance of plants is correlated due to the presence of secondary metabolites. Our plant of interest, alcohols . A novelrophorus –10.P. hysterophorus were screened for in vitro selective enzyme inhibition assays including urease, α-glucosidase, and phosphodiesterase. The ethyl acetate (87.3%), butanol (84%), and aqueous (81.4%) fractions of P. hysterophorus exhibited maximum inhibition of urease enzyme compared to other fractions, however, exhibited less activity than the standard acetohydroxamic acid (96.3%). Urease is a metalloenzyme found in bacteria and plants. The urease inhibitors are important in the conditions associated with ureolytic bacteria [H pylori infections [α-glucosidase inhibition by various fractions of P. hysterophorus highlighted its importance for controlling hyperglycemia in diabetic patients [The extracts and fractions of bacteria . Furtherfections . In agrifections . Furtherpatients , 25.P. hysterophorus is worth noting. Phosphodiesterase (PDE) has more than 40 isoforms, further subdivided into eleven families. These enzymes (present in each cell) hydrolyze the intercellular second messengers, cyclic nucleotide adenosine-3′, 5′- cyclic monophosphate (cAMP), and guanosine-3′, 5′-cyclic monophosphate (cGMP), thus altering cell response [The phosphodiesterase inhibition by the fractions of response . The PDEresponse . The PDEresponse .P. hysterophorus extracts and fractions [P. hysterophorus, whereas aqueous, chloroform, and n-hexane fractions exhibited moderate analgesic activity. Therefore, the phytochemical constituents of P. hysterophorus may play a role in inhibiting the release of pain-stimulating mediators such as prostaglandins (PGs), bradykinin, and histamine [The acetic acid-induced writhing test is used to examine the preliminary analgesic properties of ractions . The aceractions , 29. Ouristamine , 31. Amoin vivo assessment of the skeleton muscle relaxation potential of substances [The traction and chimney screening tests are common tools for the bstances . In our bstances . It is aα-glucosidase, and phosphodiesterase by its phytochemical constituents. The sedative, anti-inflammatory, and analgesic potential of plants are also validated experimentally. Based on our data, we can conclude that P. hysterophorus could be a potential source of new, less toxic, safe, and more effective candidate drugs for pharmaceutical industries, thereby decreasing the economic burden for the therapy of different diseases involving the targets such as urease, α-glucosidase, and phosphodiesterase.It is concluded that crude extracts and various fractions showed the presence of active phytochemicals such as steroids, alkaloids, and terpenoids. The polar extracts and fractions exhibited excellent enzyme inhibition potency, thereby signifying the inhibition potential of urease,


Type-2 diabetes (T2D), chronic kidney disease, and heart failure (HF) share epidemiological and pathophysiological features. Although their prevalence was described, there is limited contemporary, high-resolution, epidemiological data regarding the overlap among them. We aimed to describe the epidemiological intersections between T2D, HF, and kidney dysfunction in an entire database, overall and by age and sex.2 were defined as within the diabetes-cardio-renal (DCR) spectrum.This is a cross-sectional analysis of adults ≥ 25 years, registered in 2019 at Maccabi Healthcare Services, a large healthcare maintenance organization in Israel. Collected data included sex, age, presence of T2D or HF, and last estimated glomerular filtration rate (eGFR) in the past two years. Subjects with T2D, HF, or eGFR < 60 mL/min/1.73 m2, and 11,605 (0.84%) with HF. Overall, 12.6% had at least one condition within the DCR spectrum, 2.0% had at least two, and 0.23% had all three. Cardiorenal syndrome (both HF and eGFR < 60 mL/min/1.73m2) was prevalent in 0.40% of the entire population and in 2.3% of those with T2D. In patients with both HF and T2D, 55.2% had eGFR < 60 mL/min/1.73m2 and 15.8% had eGFR < 30 mL/min/1.73m2. Amongst those within the DCR spectrum, T2D was prominent in younger participants, but was gradually replaced by HF and eGFR < 60 mL/min/1.73m2 with increasing age. The congruence between all three conditions increased with age.Overall, 1,389,604 subjects  were included; 445,477 (32.1%) were 25– < 40 years, 468,273 (33.7%) were 40– < 55 years, and 475,854 (34.2%) were ≥ 55 years old. eGFR measurements were available in 74.7% of the participants and in over 97% of those with T2D or HF. eGFR availability increased in older age groups. There were 140,636 (10.1%) patients with T2D, 54,187 (3.9%) with eGFR < 60 mL/min/1.73mThis large, broad-based study provides a contemporary, high-resolution prevalence of the DCR spectrum and its components. The results highlight differences in dominance and degree of congruence between T2D, HF, and kidney dysfunction across ages.The online version contains supplementary material available at 10.1186/s12933-022-01521-9. Globally, approximately 537 million subjects have type-2 diabetes , 697 milFor several years, treatment options for type-2 diabetes, CKD, and HF did not necessarily overlap. For example, thiazolidinediones benefit glycemic control in patients with type-2 diabetes; and endothelin receptor antagonists improve kidney outcomes in patients with CKD. Yet, both were associated with increased risk for HF , 10. RenWe performed a cross-sectional analysis of adults registered at Maccabi Healthcare Services (MHS) in Israel. The database enjoys very high yearly retention (99%), validated type-2 diabetes and HF registries , 24, andThis is a cross-sectional and contemporary analysis of the MHS database, Israel’s second-largest healthcare maintenance organization (HMO). It was designed as a broad-based analysis of an entire database to limit population selection biases , and inclusion in type-2 diabetes or HF registry before December 31, 2019 , 24.The purpose of the MHS registries is to benefit disease management at single-physician levels and in the entire HMO scale. Inclusion in the diabetes registry was based on a validated algorithm , 26. Thi2 (CKD stage 3 or worse). The cardiorenal syndrome was defined as having both HF and eGFR < 60 mL/min/1.73 m2.Creatinine was measured in MHS central laboratories by compensated Jaffe methodology. Only measurements in out-patient settings were used to reduce biases associated with acute in-patient admissions. eGFR was calculated by the CKD-EPI Equation . The dia2, in accordance with the KDIGO classification.Age categories were used as follows: 25- < 40; 40- < 55; 55- < 65; 65- < 75; 75- < 85 and 85 + years. When required, the 25- < 40 and 40- < 55 years group were merged. The following eGFR categories were used > 90; 60- < 90; 30- < 60 or < 30 mL/min/1.73 mThis is a descriptive analysis, and no formal hypotheses were tested. Presented are numbers or percentages of patients, as specifically indicated. We did not perform comparative tests among groups. Distribution within eGFR groups is calculated out of the total relevant population as stated, including those without measurement, to reduce biases associated with lower eGFR availability in healthier populations. Where expressly indicated, the eGFR distribution is calculated only of those with available eGFR values.2, and 11,605 (0.84%) patients with HF. Prevalence of all three conditions was higher in older age groups; while type-2 diabetes prevalence trajectory mainly increased at 40- < 75 years and plateaued in older age groups, HF and eGFR < 60 mL/min/1.73 m2 prevalence was markedly higher in those older than 65 years  aged 25- < 40 years, another third (33.7%) were 40- < 55 years old, and the rest were 55 years or older (Table 2), 1.99%  had at least two conditions, and 0.23% (n = 3329) had all three  had at least one component of the DCR spectrum  and 2.30% of those with type-2 diabetes. The prevalence of cardiorenal syndrome increased with age and male sex, reaching 7.27% of participants older than 85 years , and HF were 10.12%, 3.90%, and 0.84%, respectively. Overall, 12.63% had at least one of these conditions ; 2.22% had at least two conditions, and 0.23% had all three; 0.40% had the cardiorenal syndrome. Of those within the DCR spectrum, the relative portion of type-2 diabetes gradually reduced with older age, accompanied by a relative increase in HF and CKD stage 3–5. The degree of congruence between conditions increased with age. Overall, the analysis provides a comprehensive broad-based description of the epidemiological intersections between components of the DCR spectrum across age and sex groups.This contemporary 2019 cross-sectional descriptive study presents the association between type-2 diabetes, HF, and kidney function in 1.4 million Israeli adults. Amongst the entire cohort, the prevalence of type-2 diabetes, eGFR < 60 mL/min/1.73 mThe prevalence of diagnosed diabetes, HF, or CKD is highly variable among countries and regions. It depends on population age, lifestyle and culture, genetic variations, comorbidities, documentation quality, survival differences, and availability of screening programs , 26, 28.Type-2 diabetes, CKD, and HF are components of one spectrum, which we collectively termed diabetes-cardio-renal (DCR) spectrum , 8, 22. SGLT2i were shown to improve the prognosis of all three conditions across disease stages and phenotypes –20. EarlFor many years, resources were invested in developing new drugs that target single components of the DCR spectrum, resulting in a wide array of medications that improve glycemia, reduce HF-associated risk, or have some degree of kidney protection. More recently, however, increasing efforts have tried to find drugs and especially combination therapies that may benefit specific groups of patients within the DCR spectrum –39. Furt2, without considering other markers of heart or kidney disease. While some eGFR measurements are lacking, this group includes mostly younger and healthier populations. To address this issue, we presented data both including as well as disregarding the missing eGFR values.This study includes data from one HMO; therefore, the external validity of the findings to other populations, especially non-Caucasians, is limited. However, Israel’s population has relatively high genetic variability due to large communities of immigrants and their descendants. The analysis is solely descriptive due to its cross-sectional nature; thus, it does not allow to conclude regarding causality. The retrospective nature of this analysis, and its reliance on registries, may result in diagnoses misidentification in some cases. Many relevant variables such as body mass index, HbA1c, albuminuria status, ejection fraction, HF etiology or severity, prediabetes, diabetes retinopathy or other comorbidities, and medications were not included. Accordingly, the definition of cardiorenal syndrome relies on the coexistence of HF ICD-9 diagnosis and eGFR < 60 mL/min/1.73 mThis huge, broad-based study of 1.4 million adults provides a contemporary 2019, high-resolution prevalence of the different components of the diabetes-cardio-renal spectrum. The findings highlight differences in the degree of congruence among type-2 diabetes, HF, and kidney dysfunction across age and sex groups. The results may assist in estimating patients’ and populations’ probability of having one or more components of the DCR spectrum, with practical implications for policymaking, clinical research, and daily patient care.Additional file 1: Figure S1. CONSORT diagram of subjects included in this analysis. Figure S2. Distribution by age of T2D, heart failure, and eGFR<60 mL/min/1.73 m2. Figure S3. Prevalence of T2D in patients with heart failure or eGFR<60 mL/min/1.73 m2 overall and at different age groups. Figure S4. Prevalence of heart failure by age, presence of T2D and kidney functions—in female and male participants separately. Figure S5. Distribution of patients within different kidney functions categories by presence of T2D, heart failure, or neither. Figure S6. Distribution of kidney functions in patients with heart failure by the presence of T2D.


For the Mn2+ analog, light irradiation changes the spin topology from a single Mn2+ ion to a radical-Mn2+ single chain, further inducing magnetic bistability with a remarkably wide thermal hysteresis of 177 K. Structural analysis of light irradiated crystals at 300 and 50 K reveals that the rotation of the anthracene rings changes the Mn1–O2–C8 angle and coordination geometries of the Mn2+ center, resulting in magnetic bistability with this wide thermal hysteresis. This work provides a strategy for constructing molecular magnets with large thermal hysteresis via electron transfer photochromism.Achieving magnetic bistability with large thermal hysteresis is still a formidable challenge in material science. Here we synthesize a series of isostructural chain complexes using 9,10-anthracene dicarboxylic acid as a photoactive component. The electron transfer photochromic Mn Achieving magnetic bistability with large thermal hysteresis is still a challenge in material science. Here, the authors report a Mn(II) chain complex that enables light-induced magnetic bistability with a 177 K thermal hysteresis loop. Magnetically bistable phenomena with thermal hysteresis have been observed in spin-crossover materials8, metal-to-metal charge transfer compounds11, valence tautomeric complexes13, reversible dimerization of organic π-radicals16, and complexes with dynamic coordination environments19. Although bistable systems have been extensively studied, there are still some challenges. There is a need to explore wide thermal hysteresis and develop new mechanisms for constructing magnetically bistable complexes for both fundamental research and practical applications.Magnetically bistable multifunctional materials attracted significant attention owing to their potential applications in switches, sensors, and information storageT > 100 K)24. To achieve the broad thermal hysteresis, strengthening the cooperative interactions between magnetic molecules and extending dimensionality by π–π stacking and hydrogen bonding interactions are becoming a promising strategy32. Besides the widespread spin-crossover and metal-to-metal charge transfer compounds, valence tautomeric complexes with radicals or reversible dimerization of organic π-radicals provide an effective way for magnetic bistability with relatively wide thermal hysteresis loops (ΔT > 100 K). Thus, introducing radicals into molecular magnets may be a good technique for designing systems with wide thermal hysteresis loops.The width of thermal hysteresis is vital in bistable materials. Until now, only five complexes have shown large and reproducible thermal hysteresis loops with repeated scanning  light irradiation (ca. 300–800 nm), stable radical analogs are generated via electron transfer and can provoke magnetic interactions with paramagnetic metal centers, further intriguing amazing magnetic behavior, such as photodemagnetization, magnetic transitions, and single-molecule magnetism44. Furthermore, such room-temperature photochromism provides an opportunity to realize the on/off switch of photomagnetic properties with color changes. According to recent reports, anthracene-based ligands exhibit radical-actuated photochromic behavior, in which ligand with π-conjugations can kinetically stabilize the photogenerated radicals46, and magnetic interactions operate between radicals and paramagnetic metal ions. However, magnetic thermal hysteresis is still challenging in electron transfer photochromic materials.To date, direct syntheses of stable radicals are time-consuming and include unsafe redox reactions triggered by chemical and electrochemical reactions2ADC) can act as a photoactive ligand. The reaction of H2ADC with transition metal ions yields a series of isostructural single-chain complexes of [M(ADC)(H2O)2(DMF)2]n . Photochromism is not observed in 3 and 4, whereas 1 and 2 show visible photochromic phenomena at ambient conditions due to the generation of stable diradicals after Xenon (Xe)-lamp illumination. Originated from the variations of magnetic coupling between Mn2+ ions and photogenerated radicals, compound 1 exhibits a remarkable photomagnetic response after light irradiation, resulting in an unexpectedly wide thermal hysteresis with a temperature width of 177 K . We realize a magnetic thermal hysteresis loop in radical-actuated photochromic materials, and the hysteresis loop of 177 K is significantly wide compared to those reported for reproducibly bistable magnetic systems24.In this work, we report that 9,10-anthracene dicarboxylic acid  by Shape 2.0, Supplementary Table 2+ ions. The ADC bridges two Mn2+ ions to construct a one-dimensional chain , color changes from colorless to pink and white to yellow are observed in the crystalline and powder samples, respectively . The time-dependent solid UV–Vis spectra for the samples show a broad absorption around 475 nm upon irradiation by the Xe-lamp  of ~9.20 min . Compared with 1, the photochromic rate for 2 with t1/2 of 12.31 min  spectra for 1 and 2 before and after the light irradiation remained unchanged, indicating no isomerization or/and photodimerization  was measured for all the compounds, and the experimental curves are in good agreement with the simulated one  for 2, the obtained spin concentration of 2a is clearly smaller. This discrepancy is because photochromic reactions mainly occur at material surfaces. Owing to the similarity in the similar photochromic behavior, the spin concentration of 1a should also be smaller than the theoretical value51. Thus, crystal samples of both compounds were ground to powder, and the light-irradiation time for photochromic experiments was increased to at least 180 min to ensure the sufficiently photogenerated radicals. Magnetic susceptibility measurements of 2a revealed the formation of radicals  measurements for 059 Fig. , attribu004 Fig.  after li2ADC ligand was examined under the same conditions. H2ADC shows color changes from yellow to deep yellow after Xe-lamp illumination  calculations, and a method with the basis set B3LYP/6-311G(d) using the Gaussian 09 program was performed to calculate the spatial distributions of the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) levels. As shown in Fig. 1 is mainly originated from the photoactive H2ADC ligands. However, the electron distribution in the HOMO is dominantly located on the carboxyl groups, whereas that in LUMO is dominant on the anthracene rings. The localization of HOMO and LUMO suggests the HOMO–LUMO transition is attributed to intraligand electron transfer from carboxyl groups to anthracene motifs56. For light irradiated 1a, DFT calculations also revealed that electron distribution is mainly located on the ADC ligands  value for the as-prepared sample of 1 is 4.69 cm3 K mol−1 at room temperature  with g = 2.0. As the temperature decreases, χT decreases slightly to 3.59 cm3 K mol−1 at 2 K. A Curie-Weiss plot gives a negative Weiss constant θ of −0.54 K , showing a thermal hysteresis loop with a large width of 177 K  among the reported magnetically bistable molecular systems24. The magnetic susceptibility data for 1a follows the Curie-Weiss trend  and then flipped the spins on all the atoms except for Mn2+ to obtain the energy of the low-spin (LS) state (SLS = –1/2 –1/2 + 5/2 = 3/2). The spin-density distributions 2(DMF)2 around the other ADC• is omitted. The corresponding spins on Mn2+ and one ADC• are 4.896 and 2.019 in the HS state and 4.889 and −1.970 in the LS state, respectively. Thus, only the exchange coupling between Mn2+ and one ADC• should be considered for the created fragment. The spins of Mn2+ and one ADC• are 5/2 and 1, respectively. According to Hamiltonian, 2+–ADC• coupling constant JMn–ADC was obtained based on Eqs. 63.JMn–ADC calculated using Eqs. −1, respectively. The negative JMn–ADC values suggest exchange couplings between Mn2+ and ADC• are antiferromagnetic for the 1D chain of 1a, constant with the Curie-Weiss fitting results.Furthermore, the exchange coupling constant DC• Fig.  from theons Fig.  show tha8, metal-to-metal/ligand electron transfer13, organic radicals with intra- or intermolecular electron-exchange interactions64, spin-Peierls-type transition65, transition metal complexes with dynamic coordination environments67, and angular-momentum quenching of Co2+ complexes70. However, the light-induced thermal hysteresis of 1 originates from the variations of magnetic couplings and cooperativity, which is distinct from the reported examples. To further explore the magnetic behavior of 1a, variable-temperature ESR spectra were conducted between 300 and 100 K for 1, 1a, and 2a. As shown in Fig. 1a remain unchanged, while the intensity gradually increases upon cooling, similar to the ESR variation for 1 before irradiation ° between the rings at high and low temperatures , 91.65(8) and 88.27(9) to 89.60(5)°, 92.33(4)° and 85.38(5)° upon cooling to 50 K, respectively. Thus, the geometrical configuration of the Mn2+ center suffers a more and abnormal distortion in the low temperatures , H2ADC , DMF (1.0 mL), and H2O (4.0 mL) were sealed in a glass vial and heated to 90 °C for 5 days. Colorless X-ray-quality crystals were formed and dried in air. Yield: 0.049 g (49%) based on MnCl2·4H2O. Compound 1a was obtained by 250-W Xe-lamp irradiation of 1 at room temperature for 180 min. Elemental analysis (%): calcd for 1 (%): calcd for C22H26N2MnO8 (501.39): C, 52.70; H, 5.23; N, 5.59. Found: C, 52.62; H, 5.11; N, 5.54. IR : 3515(w), 2921(w), 1656(s), 1569(m), 1434(m), 1328(w), 1276(w), 1099(w), 1028(w), 861(w), 790(m), 671(m), and 467(w). For 1a (%): calcd for C22H26N2MnO8 (501.39): C, 52.70; H, 5.23; N, 5.59. Found: C, 52.57; H, 5.31; N, 5.66.A mixture of MnCl1 by using Zn(NO3)2·6H2O . Yield: 0.045 g (44%) based on Zn(NO3)2·6H2O. Compound 2a was obtained by 250-W Xe-lamp irradiation of 2 at room temperature for 180 min. Elemental analysis (%): calcd for 2 (%): calcd for C22H26N2ZnO8 (511.82): C, 51.62; H, 5.12; N, 5.47. Found: C, 51.72; H, 4.95; N, 5.54. IR : 3567(w), 3436(w), 2931(s), 1654(s), 1554(s), 1488(w), 1432(m), 1374(w), 1322(m), 1276(w), 1245(w), 1099(m), 861(w), 782(m), 676(m), 479(w). For 2a (%): calcd for C22H26N2ZnO8 (511.82): C, 51.62; H, 5.12; N, 5.47. Found: C, 51.81; H, 5.03; N, 5.43.The crystals were prepared in a similar way with compound 1 by using Ni(NO3)2·6H2O . Yield: 0.048 g (47%) based on Ni(NO3)2·6H2O. Elemental analysis (%): calcd for C22H26N2NiO8 (505.16): C, 52.31; H, 5.19; N, 5.55. Found: C, 52.32; H, 5.08; N, 5.34. IR : 3598(w), 3430(m), 2976(s), 2929(s), 1646(s), 1550(m), 1436(m), 1374(w), 1326(w), 1272(w), 1101(w), 1051(w), 871(w), 782(m), 678(m), 584(w) 484(w).The crystals were prepared in a similar way with compound 1 by using Co(NO3)2·6H2O . Yield: 0.040 g (40%) based on Co(NO3)2·6H2O. Elemental analysis (%): calcd for C22H26N2CoO8 (508.38): C, 52.28; H, 5.19; N, 5.54. Found: C, 52.37; H, 5.28; N, 5.44. IR : 3561(w), 2933(s), 1644(s), 1552(m), 1439(m), 1376(w), 1318(m), 1276(w), 1099(w), 780(m), 671(m), 605(w), and 479(w).The crystals were prepared in a similar way with compound 2ADC ligand, and a Bruker E500 spectrometer with continues-wave X band frequencies of 9.84 GHz at room temperature and 9.41 GHz at low temperatures for 1 and 2. Magnetic measurements of the polycrystalline samples were carried out on a Quantum Design SQUID PPMS magnetometer. PXRD curves were performed on a Rigaku diffractometer with a Cu-target tube and a graphite monochromator. Simulation of the PXRD curves were performed by the single-crystal data and diffraction-crystal module of the Mercury (Hg) program available free of charge via the Internet at http://www.iucr.org. Furthermore, a Rietveld refinement of PXRD between the experimental pattern and the single crystal data were also performed. For the light irradiation experiments, a Perfect Light PLS-SXE 300 Xe-lamp  was equipped to prepare the colored samples of 1 and 2 for elemental analyses, IR, crystal XRD, UV–Vis, PXRD, ESR and magnetic studies.Elemental analyses were performed on a PerkinElmer 240C analyzer. The UV–Vis spectra were applied on a Puxi Tu-1901 spectrophotometer. The luminescence curves were recorded on a Hitachi F-7000 Fluorescence spectrometer. IR curves were collected on an ABB Bomen MB 102 series FT−IR spectrometer. The ESR spectra were measured on CIQTEK EPR200-Plus with a continues-wave X band frequency of 9.84 GHz for HJ between photogenerated radicals and Mn2+ ions was calculated by DFT calculations using the popular hybrid functional O3LYP with ORCA 5.0.2.Molecular orbital calculations were performed using the Gaussian 09 program and the basis set B3LYP/6-311G(d) method and adapted from the crystal X-ray data. The exchange coupling constant 1a with different temperatures were collected on a Bruker D8 Venture CMOS-based single-crystal X-ray diffractometer equipped with a graphite-monochromated Mo-Kα radiation (λ = 0.71073 Å) using the SMART and SAINT programs. 1–4 were collected on a Rigaku SCX-mini diffractometer with Mo-Kα radiation (λ = 0.71073 Å) at room temperature. The SHELX-2016 software was used to solve all those structures75.The single-crystal X-ray diffraction data of Supplementary InformationPeer Review File


Striga spp., and guiding them towards host plants in their vicinity. Based on their structure, SLs are divided into canonical and non-canonical SLs. In this perspective, we describe the metabolism of root-released SLs and SL pattern in rice max1-900 mutants, which are affected in the biosynthesis of canonical SLs, and show the accumulation of two putative non-canonical SLs, CL+30 and CL+14. Using max1-900 and SL-deficient d17 rice mutants, we further investigated the metabolism of non-canonical SLs and their possible biological roles. Our results show that the presence and further metabolism of canonical and non-canonical SLs are particularly important for their role in rhizospheric interactions, such as that with root parasitic plants. Hence, we proposed that the root-released SLs are mainly responsible for rhizospheric communications and have low impact on plant architecture, which makes targeted manipulation of root-released SLs an option for rhizospheric engineering.Strigolactones (SLs) are a plant hormone regulating different processes in plant development and adjusting plant’s architecture to nutrition availability. Moreover, SLs are released by plants to communicate with beneficial fungi in the rhizosphere where they are, however, abused as chemical cues inducing seed germination of root parasitic weeds, e.g.  We showed that oxo-CL, together with CL+30, are mobile SLs, as they were the only SLs detected in the shoot base (1 cm-long stem tissue corresponding to the area connecting the roots and the shoot) (d17 and d10 with the exudates collected from wild-type (WT) and max1-900 mutants. We observed interesting SL metabolic conversions as well as changes in the capability to induce parasitic weed germination. Based on our results, we propose that root-released SLs function as regulatory cues in rhizosphere communications and hypothesize CL-derived molecules might regulate rice plant architectures, although their biosynthetic enzyme(s) are yet uncharacterized.As SL biosynthesis and exudation are generally induced under nutrient-limited conditions, secreted SLs might serve as communicators between non-parasitic plants themselves as well as with other organisms in the rhizosphere. This plant-plant communication may provide an advantage to the secreting plant, allowing the occupation of the space they need to increase the chance of survival. To address this hypothesis, we re-investigated the SL profiles of the rice e shoot) . MoreoveOs900 KO-mutant lines do not contain the canonical SLs 4DO and orobanchol, but accumulate a putative 4-oxo-hydroxyl-carlactone, called CL+30  analysis. Our results confirmed the absence of 4DO and orobanchol in the Os900-KO lines, along with the accumulation of CL+30 (tentative namely 4-oxo-hydroxyl-carlactone) and the reduction in the putative 4-oxo-MeCLA , in comparison to WT (m/z 97.02 and a parent ion at m/z 317.17 in root tissues and exudates of both max1-900 lines (Os900-32 and -34) . To classify CL+14 as a SL and confirm that CL was its precursor, we fed Os900-34 rice seedlings with [13C]-labeled CL. We identified [13C]-CL+14 in the root exudates, indicating this that metabolite is a downstream product of CL  as orobanchol  with Os900-32 and -34 root exudates. Orobanchol isomer was only detected by LC-MS/MS analysis when d10 seedlings were fed with Os900 KO-lines root exudates  as a common intermediate in SL biosynthesis , the “re mutants . In thismulation . Thus, td17 feeding experiment showed that 97% of 4DO was absorbed by plants in the hydroponic system, while the uptake of orobanchol occurred at much lower level  through the action of MAX1-900 and/or other CYPs  in determining tillers number and plant’s height. Nevertheless, our data indicate that SLs released by plants do not have substantial effect on shoot growth and development. On the other hand, we demonstrate here that released SLs are important rhizosphere metabolites, and that their metabolism reduced the root exudates’ activity in inducing root parasitic seed germination. Hence, we hypothesize that rice plants have evolved metabolic pathways to modify particular SL structures for beneficial purpose, such as AM symbiosis. In addition, we propose an approach to investigate SL metabolism and probably SL signaling, by using root exudates as a feeding stock. Indeed, application of naturally accumulated SLs from SL-related mutants can be an alternative to treatment with SL chemically synthesized, expensive and in many cases commercially not vailable SLs.Finally, the effect of a SL varies based on the stereochemistry and composition of SLs and is highly species-dependent; hence, feeding with root exudates can be a useful tool to investigate SL metabolism in different plant species and to depict similarities and differences in this process. For instance, it would very interesting to check if CL-derivatives discovered in rice can be metabolized by tomato plants? Or if 4DO can be metabolized by sorghum? If so, would these specialized metabolites serve as communication signals between plants? And how do plants distinguish SLs released by their neighbors from their own?max1-900, named here Os900 (d17 (−2 s−1 and day/night temperature of 27/25°C). Rice seeds were surface-sterilized in a 50% sodium hypochlorite solution with 0.01% Tween-20 for 15 min. Seeds were then rinsed with sterile water and germinated in the dark overnight. The pre-germinated seeds were transferred to Petri dishes containing half-strength liquid Murashige and Skoog (MS) medium and incubated in a growth chamber for 7 days. Thereafter, the seedlings were transferred into black falcon tubes filled with low-Pi of half-strength modified Hoagland nutrient solution with adjusted pH to 5.8. The nutrient solution consisted of 5.6 mM NH4NO3, 0.8 mM MgSO4·7H2O, 0.8 mM K2SO4, 0.18 mM FeSO4·7H2O, 0.18 mM Na2EDTA·2H2O, 1.6 mM CaCl2·2H2O, 0.8 mM KNO3, 0.023 mM H3BO3, 0.0045 mM MnCl2·4H2O, 0.0003 mM CuSO4·5H2O, 0.0015 mM ZnCl2, 0.0001 mM Na2MoO4·2H2O and 0.004 mM K2HPO4·2H2O.re Os900 , d17  preconditioned with 3 mL of methanol and 3 mL of water. After washing with 3 mL of water, SLs were eluted with 5 mL of acetone. The SL fraction was concentrated to SL aqueous solution (∼1 mL), followed by 1 mL of ethyl acetate extraction. 750 μL of SL enriched organic phase was dried under vacuum. For root tissues extraction, we followed the protocol described in  for 15 min, followed by centrifugation for 8 min at 3800 rpm at 4°C. The two supernatants were combined and dried under vacuum. The residue was dissolved in 50 μL of ethyl acetate and 2 mL of hexane, following a Silica Cartridges SPE column (500 mg 3 mL-1) purification. After washing with 3 mL of hexane, SLs were eluted in 3 mL of ethyl acetate and evaporated to dryness under vacuum. The final extract was re-dissolved in 100 μL of acetonitrile: water  and filtered through a 0.22 μm filter for LC-MS/MS analysis.SLs in rice root exudates were identified based on the protocol published in . In shorribed in . Around 18 Selectivity HPLC Columns  with mobile phases consisting of water (A) and acetonitrile (B), both containing 0.1% formic acid, and the following linear gradient : 0–15 min, 25%–100% B, followed by washing with 100% B and equilibration with 25% B for 3 min. The injection volume was 10 μL, and the column temperature was maintained at 30 °C for each run. The MS conditions were as follows: positive mode, ion source of H-ESI, spray voltage of 3500V, sheath gas flow rate of 60 arbitrary units, auxiliary gas flow rate of 15 arbitrary units, sweep gas flow rate of 2 arbitrary units, ion transfer tube temperature of 350 °C, vaporizer temperature of 400 °C, S-lens RF level of 60, resolution of 120000 for MS; stepped HCD collision energies of 10 to 50% and resolution of 30000 for MS/MS. The mass accuracy of identified compounds (accurate mass ± 5 ppm mass tolerance) were acquired using Xcalibur software version 4.1.The identification of SLs was performed by using UHPLC-Orbitrap ID-X Tribrid Mass Spectrometer (Thermo Scientific™) with a heated-electrospray ionization source. Chromatographic separation was achieved on the Hypersil GOLD C6-5-deoxystrigol. 299.09→158.06, 299.09→157.06, 299.09→97.02 for GR24. 317.17→ 220.14, 317.17→205.12, 317.17→164.08, 317.17→97.02 for CL+14 (putative oxo-CL).SLs were quantified by LC-MS/MS using a UHPLC- Triple-Stage Quadrupole Mass Spectrometer (Thermo Scientific™ Altis™). Chromatographic separation was the same as above SL identification. The MS parameters were: positive ion mode, ion source of H-ESI, ion spray voltage of 5000 V, sheath gas of 40 arbitrary units, aux gas of 15 arbitrary units, sweep gas of 2 arbitrary units, ion transfer tube gas temperature of 350 °C, vaporizer temperature of 350 °C, collision energy of 17 eV, CID gas of 2 mTorr. The characteristic Multiple Reaction Monitoring (MRM) transitions (precursor ion → product ion) were 331.15→216.0, 331.15→234.1, 331.15→97.02 for 4-deoxyorobanchol; 347.14→329.14, 347.14→233.12, 347.14→ 205.12, 347.14→97.02 for orobanchol; 361.16→ 247.12, 361.16→177.05, 361.16→208.07, 361.16→97.02 for putative 4-oxo-MeCLA; 333.17→219.2, 333.17→173.2, 333.17→201.2, 333.17→97.02 for CL+30 (putative 4-oxo-hydroxyl-CL); 337.19→222.15, 337.19→240.16, 337.19→97.02 for DC-CL was prepared following the protocol described previously  containing the root exudates of two-weeks old WT, Os900-32 or -34 plants thatwere hydroponically grown two plants per 50 mL black tube under low Pi conditions. Pi content and pH of collected exudates were readjusted to 5.8 by the addition of K2HPO4.2H2O and HCl, before using them for feeding. Fresh low Pi solution was provided to the WT, Os900-34 and -34 seedlings daily; 6 h later, the exudates from the same genotype were pooled together, adjusted (Pi concentration was 0.4 mM) and fed to d17 for two week.Os900-34 and -34 seedlings daily; 24 h later, the exudates from the same genotype were pooled together, adjusted (Pi concentration was 0.4 mM) and fed to two week Pi starvation d17 and d10 seedlings for 6 h incubation; thereafter exudates were extracted according to d mutants)-(unfed exudate)/unfed exudate]*100For SL analysis, fresh low Pi solution was provided to the WT, 2O), 5 μL 2X diluted SYBR Green Reaction Mix, and 0.5 μL of each primer (2 μM working stock). All reactions were performed on a 384-well plate in a CFX384 Touch Real-Time PCR Detection System (Bio-Rad) as follows: 95°C for 90 sec, 40 cycles of 95°C for 15 sec, 60°C for 30 sec. All reactions were performed on at least three biological and three technical replicates, including a water control to exclude potential unspecific amplification. The 2-ΔΔCT method was used to calculate the relative gene expression levels , according to the manufacturer’s instructions. cDNA was synthetized from 2 µg of total RNA using iScript cDNA Synthesis Kit  according to the instructions in the user manual. qRT-PCR was performed using SYBR Green Master Mix (Applied Biosystems), which was diluted before use. Each PCR reaction was carried out in a total volume of 10 μL containing 4 μL diluted cDNA  and non-germinated seeds were counted under a binocular microscope to calculate germination rate (%) by using the open-source SeedQuant software . All statistical elaborations were performed using GraphPad Prism, version 8.3.0.Data are represented as mean and their variations as standard deviation. The statistical significance was determined by one-way analysis of variance (one-way ANOVA) with Tukey’s multiple comparison test, using a probability level of p<0.05; or two-tail student t-test with denote significant differences  given to SA-B.Striga hermonthica seeds.We sincerely thank the support from the members of KAUST Analytical Core Lab and the Bioactives lab. We thank Dr. Abdel Gabbar Babiker for providing The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Acidification can seriously affect the growth of tea trees and the yield and quality of tea leaves. In this study, we analyzed the effects of acidification on the physicochemical properties, microorganisms and metabolites of tea rhizosphere soils with different pH values, and the results showed that with the increase of soil pH, the organic matter content, cation exchange capacity, microbial biomass carbon, microbial biomass nitrogen, microbial respiration intensity, bacterial number and actinomyces number in tea rhizosphere soil all showed an increasing trend, while the fungi number decreased. The results of soil metabolite analysis showed that 2376, 2377 and 2359 metabolites were detected in tea rhizosphere soil with pH values of 3.29, 4.74 and 5.32, respectively, and the number of similar compounds reached 2331, accounting for more than 98%. The results of soil metabolite content analysis showed that with the increase of soil pH, the total contents of metabolite of tea rhizosphere soil increased significantly. The results of correlation analysis between physicochemical indexes of soil and microorganisms and soil metabolites showed that physicochemical indexes of soil and microorganisms were significantly correlated with 221 soil metabolites, among which 55 were significantly positively correlated and 166 were significantly negatively correlated. Based on correlation interaction network analysis, 59 characteristic compounds were obtained and divided into 22 categories, among which 7 categories compounds showed a significant increasing trend with the increase of soil pH, while the other 15 categories compounds showed the opposite trend. Based on the functional analysis of characteristic metabolites, this study found that with the increase of soil pH in tea rhizosphere, the diversity and number of soil microorganisms increased, and the cyclic ability of C and N of tea rhizosphere soil was enhanced, which in turn might lead to the enhancement of resistance of tea tree and promote the growth of tea tree. Tieguanyin is one of the top ten famous teas in China, and Tieguanyin tea tree is native to Anxi County, Quanzhou City, Fujian Province, China . Anxi County is the main tea producing county in China, with tea production ranking first in China for 8 consecutive years (From 2014 to 2021). Tea tree is an acidophilic crop, however, when the soil pH was lower than 4.0, the soil was severely acidified and the growth of tea tree and its tea yield and quality were significantly reduced  at a soil to water ratio of 1:2.5, and 5 replicates were used for each sample . Soil or2.32/kg·h) was measured using the alkali absorption method, based on the amount of CO2 released from soil, per hour per kilogram  at a dosage of 0.5 g fresh soil. DNA was detected by 1% agarose gel electrophoresis, and purified using gel recovery kit . The purified DNA was used for quantitative 2O; the PCR reaction procedures was 94 °C pre-denaturation for 4 min, 94 °C for 1 min, 55 °C for 1 min, 72 °C for 1 min, and fluorescence signals were collected for 30 cycles.The primers used for quantitative analysis of bacterial were F27 5’-AGAGTTTGATCMTGCCTCAG-3’, R1492 5’-TACHHYTACCTTGTTACGACTT-3’. The PCR reaction system was 25 μL, including 0.5 μL of template DNA, 12.5 μL of SYBR Premix Ex Taq (TaKaRa Biotechnology), 1 μL of each primers, and the rest was ddH2O.The PCR reaction procedure was 95 °C pre-denaturation for 5 s, 94 °C for 30 s, 53 °C for 30 s, 72 °C for 30 s, fluorescence signals were collected for 40 cycles.The primers used for quantitative analysis of fungi were 5.8S 5’-CGCTGCGTTCTTCATCG-3’, ITSIF 5’-CTTGGTCATTTAGAGGAAGTAA-3’. The PCR reaction system was 25 μL, including 1 μL of template DNA, 12.5 μL of SYBR Premix Ex Taq (TaKaRa Biotechnology), 1 μL of each primer, and the rest was ddH2O; the PCR reaction procedures was, predenaturation at 95°C for 10 min, 95°C for 15 s, 65°C for 30 s, 72°C for 15 s, fluorescence signals were collected, and 40 cycles were performed.The primers used for quantitative analysis of actinomyces were Act920F 5’-TACGGCCGCAAGGCTA-3’, Act1200R 5’-TCRTCCCCACCTTCCTCCG-3’. The PCR reaction system was 25 μL, including 1 μL of template DNA, 12.5 μL of SYBR Premix Ex Taq (TaKaRa Biotechnology), 1 μL of each primer, and the rest was ddH2.52.5.1The non-targeted metabolomics method was used to analyze the metabolites in the rhizosphere soil of tea tree. This was done by taking 50 mg of soil samples and homogenizing them with 500 μL of ice-cold methanol/water , respectively. The mixtures were homogenized at 30 Hz for 2 min. After homogenization, the mixture was shaken for 5 min and incubated on ice for 15 min, and centrifuged at 12,000 rpm and 4 °C for 10 min and suck 400 μL of supernatant into another centrifuge tube. 500 mL of ethyl acetate/methanol  was added to the original centrifuge tube, oscillated for 5 min and incubated on ice for 15 min, centrifuged at 12,000 rpm and 4 °C for 10 min, and then took 400 mL of supernatant. The two supernatants were combined and concentrated. Then 100 μL of 70% methanol water was added into the dried product for 3min of ultrasonication. Finally, the product was centrifuged at 12,000 rpm and 4 °C for 3 min, and 60 μL of supernatant was aspirated for LC-MS/MS analysis.2.5.2All samples were acquired by the LC-MS/MS system followed machine instructions. Analytical conditions of UPLC were as follows: column, Waters ACQUITY UPLC HSS T3 C18 ; column temperature, 40 °C; flow rate, 0.35 mL/min; injection volume, 2 μL; solvent system, water (0.04% acetic acid): acetonitrile (0.04% acetic acid); gradient program of positive ion, 95:5 V/V at 0 min, 79:21 V/V at 3.0 min, 50:50 V/V at 5.0 min, 30:70 V/V at 9.0 min, 5:95 V/V at 10.0 min, 95:5 V/V at 14.0 min; gradient program of negative ion, 95:5 V/V at 0 min, 79:21 V/V at 3.0 min, 50:50 V/V at 5.0 min, 30:70 V/V at 9.0 min, 5:95 V/V at 10.0 min, 95:5 V/V at 14.0 min. In MS analysis, positive ion mode and negative ion mode of electrospray ion source were adopted. The positive ion conditions were: voltage 250 V, gas flow 8 L/min, fragmetor 135 V, gas temperature 325 °C, sheath temperature 325 °C, sheath flow 11 L/min, and nebulizer 40 psi. The negative ion conditions were: voltage 1500 V, gas flow 8 L/min, fragmetor 135 V, gas temperature 325 °C, sheath temperature 325 °C, sheath flow 11 L/min, and nebulizer 40 psi.2.5.3The original data file obtained by LC-MS analysis was firstly converted into mzML format by Proteowizard software. Peak extraction, alignment and retention time correction were performed by XCMS program. The “SVR” method was used to correct the peak area. Filter the peaks with deletion rate > 50% in each group of samples. After that, metabolic identification information was obtained by searching the laboratory’s self-built database and integrating the public database and metDNA.2.6Excel 2017 software was used to calculate the average value and variance of data, Rstudio 3.3 software was used to make Cloud and rain map, correlation matrix maps, violin chart, box chart, principal component maps, venn diagram, and bubble map; Heml 1.0 software was used to make heat maps; Cytoscape_v3.9.1 software was used to make the correlation analysis chart.33.12/kg·h), bacterial number (7.65 ~ 15.42 ×109 cell/g·soil), actinomyces number (2.65 ~ 7.52 ×109 cell/g·soil) all showed an increasing trend, while the number of fungi (7.43~2.48 ×109 cell/g·soil) decreased.Tea tree is an acidophilic crop, however, changes in soil pH can affect the growth of tea tree . The resThe results of correlation matrix analysis showed Figure 23.2In order to adapt to environmental changes during the growth, plants regularly modified the rhizosphere microecological environment by regulating the type or amount of their root exudes to ensure their own growth . pH valu3.3The results of the analysis of the classification of metabolites and their contents in rhizosphere soil of tea tree  can be found in the article/JY and YuhW: Conceptualization, Visualization, Methodology, Writing – original draft, Formal analysis, Writing – review & editing, Funding acquisition. SL and YucW: Formal analysis, Writing – review & editing. PC and LH: Methodology, Investigation, Writing – original draft. XJ and JK: Methodology, Investigation. ZW and HW: Conceptualization, Visualization, Methodology, Writing – original draft, Formal analysis, Writing – review & editing, Funding acquisition. All authors contributed to the article and approved the submitted version.


Brucine exhibits an inhibitory effect against Helicobacter pylori. In vivo findings revealed that brucine (3 mg/kg) showed effective results in healing ethanol-induced ulcer lesions of the gastric region in rats. Brucine showed an inhibitory effect against H+/K+-ATPase. Levels of glutathione, glutathione-s-transferase, and catalase were enhanced in the gastric rat tissue with the use of brucine, while a significant decrease in lipid peroxide levels was seen. Histopathological evaluation showed improvement in cellular architecture and a decrease in inflammatory indicators like cyclooxygenase, tumor necrosis factor, and nuclear factor kappa B expression, validated through immunohistochemistry, enzyme-linked immunosorbent assay, and Western blot techniques. In the reverse transcription–polymerase chain reaction, brucine decreased H+/K+-ATPase mRNA levels. This study reveals that brucine possesses stable binding affinities against selected targets. Brucine exhibits an anti-ulcer effect, mediated via anti-H. pylori, H+/K+-ATPase inhibition, and antioxidant and anti-inflammatory pathways.Gastric ulcer is one of the most common chronic gastrointestinal diseases characterized by a significant defect in the mucosal barrier. The current study has been conducted to evaluate the brucine anti-ulcer effect. Brucine has binding energy values ranging from −2.99 to −8.11 kcal/mol against chosen targets, according to  Gastric ulcer is a disease characterized by secondary damage produced by excessive pepsin and stomach acid output. Frequent alcohol intake has a positive link to gastric mucosal lesions that include gastritis, gastric ulcer, and in severe conditions, also gastric carcinoma . DespiteHelicobacter pylori (H. pylori) is a Gram-negative bacterium that can cause chronic gastritis, peptic ulcers, gastric adenocarcinoma, and mucosa-associated lymphoid tissue lymphoma in humans. The infection caused by this pathogen estimates about half of the world's population  and two antimicrobial drugs, amoxicillin and clarithromycin, is thought to be the most effective treatment for dication . Currentecretion . Joint decretion .The ethanol-induced gastric ulcer rodent model is considered a well-established model that reflects several features of human gastric injury and thus provides a means for testing unexplored test samples with anti-ulcer potential .Strychnos nux-vomica L. (Loganiaceae), is a weak indole alkaloid. Maqianzi, also known as Nux-vomica, has a bitter flavor . Omeprazole and metronidazole were purchased from Barrett Hodgson and Sanofi Aventis, Pakistan. Abcam, United Kingdom, provided secondary antibodies. Elabscience, China, provided the rat NFB ELISA kit , rat TNF-α ELISA kit , and rat PGE2 ELISA kit . All of the chemicals used in this experiment were of analytical quality.Sprague–Dawley rats weighing 150–200 g of either gender, purchased from the Riphah Institute of Pharmaceutical Sciences (RIPS) in Islamabad, were subjected to conduct this research work. The rats were kept in standard cages with a constant temperature of 22°C. All of the animals were fed a regular meal and had unlimited access to water. The experiments were performed by following the guidelines and principles of the Institute of Laboratory Animal Resources, Commission on Life Sciences University, National Research Council (1996). The study was approved by the Research and Ethics Committee of RIPS (Ref. No. REC/RIPS/2017/006).+/K+-ATPase (PDB ID: 5ylu), histaminergic (H2) (PDB ID: P25021), cyclooxygenase-1 (COX-1) (PDB ID: 6Y3C), tumor necrosis factor (TNF-α) (PDB ID: 1BKC), nuclear factor kappa B (NFκB) (PDB ID: 4Q3J), prostaglandin (PGE2) (PDB ID: 6AK3), cyclooxygenase-2 (COX-2) (PDB ID: 5IKQ), and muscarinic (M1) (PDB ID: 5CXV). The ligand and water molecules were removed from DSV, H-atoms (polar) were inserted, and the file was saved in PDB format. For molecular docking, the AutoDock tool-1.5.6 and PyRx-0.8 docking tools were used. The atomic contact energy (ACE)  value was calculated from the results. For post-dock analysis, the best pose with the lowest ACE value  was chosen. 2D and 3D images were assessed to determine interactions between valine (VAL), tyrosine (TYR), tryptophan (TRP), threonine (THR), serine (SER), proline (PRO), phenylalanine (PHE), isoleucine (ILE), histidine (HIS), glycine (GLY), glutamine (GLN), glutamic acid (GLU), cysteine (CYS), and aspartic acid (CYS) residues and ligand residue (ALA).The three-dimensional structure of a typical medication was obtained using a Biovia Discovery Studio Visualizer (DSV). The standard drug omeprazole was used. In gastric ulcer pathophysiology, human protein targets (3D structure) involved were selected and acquired from an online protein data bank, Research Collaboratory for Structural Bioinformatics (RCSB) PDB. The target proteins selected were HH. pylori was evaluated. From the gastric biopsy of a patient with gastric ulcer, three strains of H. pylori were received under consent at the care endoscopy clinics and laboratories . Biopsies to be analyzed were placed in a modified Campy-Thio medium. Plates were kept at 37°C in a microaerophilic environment. Identification of isolates was carried out by morphology and using a urease test kit. Isolates were kept at 80°C in sterile McCartney bottles containing 0.2 g/L cysteine and 20% glycerol in brain heart fusion broth. Frozen clinical isolates were injected onto Mueller–Hinton agar plates for subsequent inoculation. Brucine with various concentrations of 0.5, 1, 2, 4, 8, 16, and 32 µg/disc was run on standard disks and then placed on a Mueller–Hinton agar plate. Following an incubation period of 3 to 5 days at 37°C, the zone of inhibition for each disk was measured. All tests were carried out in triplicate, and the antibacterial activity was calculated as the average inhibitory diameter (mm). Metronidazole was used as a positive control . Body weight saline (10 ml/kg) was given to group I as a negative control. Group 2 was treated with ethanol at a dose of 1 ml/100 g. Groups 3 and 4 were pretreated with brucine at doses of 1 and 3 mg/kg (p.o), respectively, and group 5 received omeprazole (20 mg/kg) as a standard drug it was used. After 1 hour of all treatments, absolute ethanol (1 ml/100 g) was orally administered to each rat except the animals of group 1, which served as a negative control. All rats were sacrificed by cervical dislocation after 1 h of ethanol administration. The stomachs of the participants were separated and cleaned with normal saline before the lesion index was computed by measuring each lesion in millimeters along its greater curvature. Each lesion’s covered surface area was measured and evaluated using the method provided by 2, 2: 0.5 < US ≤ 2.5 mm2, 3: 2.5 mm2 < US ≤ 5 mm2, 4: 5 mm2 < US ≤ 10 mm2, 5: 10 mm2 < US ≤ 15 mm2, 6: 15 mm2 < US ≤ 20 mm2, 7: 20 mm2 < US ≤ 25 mm2, 8: 25 mm2 < US ≤ 30 mm2, 9: 30 mm2 < US ≤ 35 mm2, and 10: US > 35 mm2. For each stomach, the sum of the length (mm) of all lesions was used as the ulcer index (UI). The percentage inhibition (% I) was calculated usingFor gastric lesion induction, 24 h fasted rats were randomly assigned into five different groups  using a commercially available calorimetric H+/K+-ATPase screening kit having catalog no: E-BC-K122-S; Elabscience, United States. The H+/K+-ATPase activity was assessed after ATP hydrolysis by monitoring inorganic phosphate levels. The homogenate of stomach tissue was centrifuged for 10–15 min at 3,500 rpm. As a result of the supernatant, spectrophotometrically at 660 nm, the inorganic phosphate release was determined. One ATPase activity unit has been described as one µ mole of inorganic phosphorus released by ATP hydrolysis through ATPase at 1 mg per hour of tissue protein. It was then expressed as µmol pi/mg prot/hour . A microplate reader was used to measure the absorbance at a wavelength of 532 nm, where quantitative measurement of LPO was expressed in TBARS nmoles/min/mg of protein , catalase, glutathione-S-transferase (GST), and lipid peroxidation (LPO). The oxidation of GSH and DTNP produced a yellow end product, which was used to determine their levels. Using a GSH microplate reader, the absorbance of 2-nitro-5-thiobenzoic acid was determined at 412 nm. The extinction coefficient of the product produced was used to calculate the GST activity, which was reported as mole/mg of CDNB conjugate/min/mg of protein in moles/mg of protein. The synthesis of the CDNB conjugate was used to determine the GST level, and the absorbance at 340 nm was measured. In the presence of catalase, the degradation of H protein .On coated slides, in 100 percent pure xylene, tissue sections were deparaffinized and then rehydrated in 70 percent ethyl alcohol. The slides were immersed in hematoxylin for 10 min after being cleaned with distilled water. Then, in a glass jar, they were placed under running water for 10 min and treated with 1% HCl and 1% ammonia water. The slides were immersed in eosin solution for 5–10 min. After the required length of time had passed, the slides were cleaned in water and air–dried. In graded ethyl alcohol, the dried slides were dehydrated . The slides were cleaned with xylene and mounted with glass coverslips. The slides were photographed using an Olympus light microscope and analyzed by ImageJ, a computer-based application, with particular attention paid to the gastric cell size and shape, inflamed infiltrating cells, and vacuolation. The TIF images were arranged at the same threshold intensity for all groups and examined in GraphPad Prism.2O2). The slides were incubated for a period of time in a solution containing 0.1 percent Triton X-100 and 5 percent normal goat serum. After being blocked, the slides were treated overnight with murine anti-TNF-α, p-NFkB, and anti-COX-2 antibodies . Slides were handled for incubation with the biotinylated secondary antibody (dilution 1:50), depending on the source of the primary antibody, and the serum was used the next morning after cleaning with 0.1 M PBS. The ABC Elite kit was used to incubate slides in a humidified room for 1 h after secondary antibody application (Santa Cruz Biotechnology). The slides were cleaned in 0.1 M PBS, dyed with DAB solution, rinsed with distilled water, dehydrated in a graded ethanol series, settled in xylene, and mounted. A light microscope was used to capture immunohistochemical TIF pictures. By modifying the backdrop of photos according to the threshold intensity and analyzing p-NFκB, COX-2, and TNF-α positive cells at the same threshold intensity for all groups, ImageJ software was utilized to quantitatively detect hyperactivated p-NFκB, COX-2, and TNF-α. The relative integrated density of the samples compared to saline is used to compute the intensity , Rat PGE2 ELISA kit , and Rat NFκB ELISA kit , respectively, in accordance with the manufacturer’s guidelines (Elabscience). Using the Silent Crusher M (Heidolph) apparatus, the tissues were homogenized at 15,000 RPM; after centrifugation, the supernatant was collected . TNF-α, PGE2, and p-NFκB concentrations were measured using an ELISA microplate reader.  protein test kit was used to assess the protein content. On a 12 percent sodium dodecyl sulfate-polyacrylamide gel, 30 g of protein homogenate was electrophoretically separated and transferred to a polyvinylidene fluoride membrane. Membranes were incubated overnight at 4°C with primary antibodies such as p-NFκB and TNF-α and then blocked for 1 h at room temperature with 5% bovine serum albumin. After being cleansed three times with Tris-buffered saline with 0.1 percent Tween 20, the membranes were further subjected to a 1:1,000 dilution of secondary antibodies, such as goat and anti-rabbit, for 90 min at room temperature. To see the immunoreactive bands, an enhanced Western blotting substrate kit was used. Densitometry was used to assess the measurement of protein expression by ImageJ software. .+/K+-ATPase was normalized to the expression of β-actin as a house-keeping gene by the 2-∆∆CT method. Primer sequences for H+/K+-ATPase and β-actin were CCCGCGAGTACAACCTTCT (forward) and CGTCATCCATGGCGAACT (reverse) and TATGAATTGTACTCAGTGGA (forward) and TGGTCTGGTACTTCTGCT (reverse), respectively  from gastric tissues, as directed by the manufacturer. Using a reverse transcriptase enzyme mix and a PCR thermocycler, the first strand of cDNA was generated from 1 to 2 g of total RNA. The mRNA expression of Hectively .post hoc Tukey’s test. p < 0.05 was regarded as statistically significant.The data are presented as a mean with a standard error of the mean (SEM). GraphPad Prism 8 was used to apply statistical parameters such as one-way analysis of variance (ANOVA) with +/K+-ATPase, H2, COX-1, TNF-α, NFκB, PGE2, COX-2, and M1, the values of atomic energy  of ideal dock postures of brucine and reference drug, as well as residues implicated in H-bonding, pi-pi bonding, and other hydrophobic interactions, are shown. +/K+-ATPase receptor and produced no H-bonds or hydrophobic contacts. Brucine had an E-value of -8.1 kcal/mol against H2 and produced two H-bonds, two bonds, and no hydrophobic interaction. Brucine has an E-value of −7.6 kcal/mol against COX-1, forming two H-bonds and no hydrophobic contacts. Brucine had an E-value of −8.11 kcal/mol against TNF-α and produced one H-bond, no bond, and no hydrophobic interaction. Brucine had an E-value of −8.58 kcal/mol against NFκB, forming two H-bonds and no hydrophobic interactions. Brucine had an E-value of −6.92 kcal/mol against PGE2 and produced no H-bonds, one bond, and no hydrophobic bonds. Brucine has an E-value of –2.99 kcal/mol against COX-2, forming one H-bond and no hydrophobic interactions. Brucine has an E-value of −8.74 kcal/mol against M1, forming one H-bond and no hydrophobic interactions.  J63 (cag A-), strain (B) j196 (cag A-), and strain (C) j107 (cag A+). Different concentrations of brucine were used against strain (A) that are 0.5, 1, 2, 8, 16, and 32 µg/disc, and zones of inhibition were calculated as 1, 1.33, 2, 4, 8, 10.66, and 14 mm, respectively, and its MIC is 16 μg/ml. The same concentration of brucine was used for strain (B), and the inhibition values are 1.33, 1.66, 2, 4.66, 7.33, 10, and 13 mm, respectively, and its MIC is 18 μg/ml. For strain (C), same concentrations of brucine were used, and the inhibitory values are 1.66, 2, 2.33, 4.33, 8.66, 11.66, 14, and 16 mm, respectively, and its MIC is 16 μg/ml, compared with the standard metronidazole, respectively (The minimum inhibitory concentration (MIC) and zone of inhibition of three different strains of ectively .Brucine at 1 and 3 mg/kg exhibited an anti-ulcer effect. Brucine shows an 80% protective effect at 1 and 3 mg/kg doses with ulcer indexes of 2 ± 0.93 and 2 ± 0.31, respectively. Omeprazole (20 mg/kg) exhibited a 90% inhibitory effect with the ulcer index 1 ± 0.13 as compared to the ethanol group . The sto+/K+-ATPase activity was increased by 103.75 µmol Pi/mg prot/h, while brucine (3 mg/kg) and omeprazole (20 mg/kg) reduced H+/K+-ATPase by 24.34 µmol Pi/mg prot/h and 30.29 µmol Pi/mg prot/h, respectively  and omeprazole (20 mg/kg)-treated groups significantly reduced LPO while restoring GST, GSH, and catalase .Normal stomach tissues were seen in the saline (10 ml/kg) group, having architecture without any pathological changes. Ethanol (1 ml/100 g)-ulcerated rat tissues exhibited severe gastric tissues, damage with vacuolation, and disruption of morphological cell boundaries. The gastric tissues of mice treated with brucine (3 mg/kg) and omeprazole (20 mg/kg) demonstrated the regeneration and repair of gastric cells with minimal degradation .The results of IHC on stomach tissues revealed that the ethanol (1 ml/100 g)-treated group markedly upregulated inflammatory markers COX-2, p-NFκB, and TNF-α expression. Vacuolation, necrotic cells, and disruption of morphological cell boundaries were found in the disease group. Brucine (3 mg/kg) and omeprazole (20 mg/kg) groups downregulated COX-2, p-NFκB, and TNF-α expressions , 7.2 levels were 1960 ± 50, 1980 ± 30, and 1755 ± 55 pg/ml, respectively. In the ethanol (1 ml/100 g)-treated group, p-NFκB and TNF-α levels increased to 3,888 ± 88 and 3,896 ± 75 pg/ml, and the PGE2 level decreased to 785 ± 25 pg/ml, respectively. In the brucine (3 mg/kg)-treated group, p-NFĸB and TNF-α levels decreased to 2,740 ± 70 and 2,730 ± 80 pg/ml, and the PGE2 level increased to 850 ± 50 pg/ml, respectively. In the omeprazole (20 mg/kg)-treated group, p-NFκB and TNF-α levels decreased to 2,365 ± 85 and 2,365 ± 85 pg/ml, and the PGE2 level increased to 1765 ± 85 pg/ml, respectively -treated group, p-NFκB, TNF-α, and PGEectively .In the ethanol (1 ml/100 g) group, p-NFκB and TNF-α expressions in the gastric region were increased. In brucine (3 mg/kg)- and omeprazole (20 mg/kg)-treated groups, TNF-α and NFκB levels were suppressed .+/K+-ATPase in ethanol-induced gastric ulcer. In the ethanol-treated group, the expression of H+/K+-ATPase mRNA levels increased. Brucine (3 mg/kg) and omeprazole (20 mg/kg) decreased H+/K+-ATPase mRNA levels (RT-PCR determined the fold expression of HA levels .1 > H+/K+-ATPase > NFκB > H2 > COX-1 > PGE2 > COX-2. This study displayed results of all ligands compared with standard drugs, obtained from the PubChem database and RCSB.This research work was conducted to explore the protective actions of brucine against ethanol-induced gastric ulcers in Sprague–Dawley rats. Ethanol is known for its notorious gastric ulceration by directly enhancing the disruption of mucosal cellular membranes, dehydration, and cytotoxicities, followed by the propagation of inflammatory cytokines, oxidative damage, and apoptosis . The curin vivo experimentation (in vivo study, pretreatment of rats with brucine at both doses significantly reduced the ulcer index relative to the disease group. In comparison to the usual treatment, ulcerated animals pretreated with brucine demonstrated a greater reduction in the ulcer index as compared to omeprazole (In the present study, the ethanol-induced gastric ulcer model was used for entation . In an ieprazole . In humaeprazole .in vitro conformational analysis, H. pylori is the main risk factor for gastric ulcer disease (H. pylori bacterium which is mainly responsible for gastric ulceration (H. pylori activity was examined through the zone of inhibition and minimum inhibitory concentration. Brucine showed an anti-H. pylori effect through the zone of inhibition and MIC against three different clinical strains.In the  disease . Brucineceration . Anti-H.+/K+-ATPase has a key role in gastric ulcer. The proton pump in the stomach and the enzyme that is responsible for acidification of the stomach contents are known as gastric H+/K+-ATPase (+/K+-ATPase, which inhibits gastric acid output, is a popular clinical intervention for dyspepsia, peptic ulcer, and gastroesophageal reflux disease. Brucine showed effective results in the inhibition of the H+/K+-ATPase enzyme (H+-ATPase . In gaste enzyme .Considerable evidence shows that inflammation is followed by oxidative stress in gastric ulcer . AccordiBrucine protected gastric cells by inhibiting inflammatory markers such as TNF-α, p-NFκB, and COX-2 when examined for cytokine levels by H&E staining and IHC, whereas these markers were significantly higher in the ethanol group. TNF-α, COX-2, and p-NFκB are prototypic pro-inflammatory cytokines because of their important role in initiating the cascade of cytokine activation and growth factors in the inflammatory response .2. These inflammatory markers were significantly raised in ulcerative tissues (For further confirmation, ELISA was carried out to quantify the inflammatory markers of TNF-α, p-NFκB, and PGE tissues , while bWestern blot findings provide evidence that brucine has anti-inflammatory effects through reduced expressions of p-NFκB and TNF-α. Both proteins are involved in the recruitment of inflammatory mediators. Western blot results show increased expressions in the ethanol group of rats compared to the saline group, while brucine and omeprazole reduced these expressions significantly . Similar+/K+-ATPase. The amounts of H+/K+-ATPase mRNA were measured in the saline, ethanol, treatment, and omeprazole groups. Results showed that H+/K+-ATPase mRNA levels increased in the ethanol group as compared to the saline group, while treatment with brucine and omeprazole significantly reduced H+/K+-ATPase mRNA levels in the treatment group as compared to the ethanol group (via anti-H. pylori, H+/K+-ATPase inhibition, and antioxidant and anti-inflammatory pathways.The RT-PCR technique provides further evidence and confirmation of targeted Hol group . It was via anti-H. pylori, H+/K+-ATPase inhibition, and antioxidant and anti-inflammatory pathways. Thus, the current findings are a reflection of the potential benefits of brucine as an effective and safer candidate for the management of gastric injuries; however, we recommend further studies to investigate in detail the potential efficacy and safety of brucine in the clinical setting as an adjunct approach to address gastric injury-related health issues.The current study highlights pieces of evidence for the protective effects of the brucine ethanol-mediated gastric injury rat model. Brucine possesses binding energy values ranging from F02D2.99 to F02D8.11 kcal/mol against selected targets. Brucine exhibits an anti-ulcer effect, mediated