Babao Dan improves neurocognitive function by inhibiting inflammation in clinical minimal hepatic encephalopathy.
BACKGROUND AND PURPOSE
Inflammation has been considered a precipitating event that contributes to neurocognitive dysfunction in minimal hepatic encephalopathy (MHE). Inhibition TLR-4 related inflammation can effectively improve neurocognitive dysfunction of MHE. Our previous study showed that Babao Dan (BBD) effectively inhibited inflammation and ameliorated neurocognitive function in rats with acute hepatic encephalopathy (HE) and chronic HE. The mechanism may lie in the regulation of TLR4 signaling pathway. Therefore, this study aimed to evaluate the role of BBD in the treatment of MHE patients with cirrhosis and to elucidate the underlying mechanism by which BBD regulated TLR4 pathway to alleviate inflammation.


METHODS
A randomized controlled trial (n = 62) was conducted to evaluate the clinical efficacy between BBD plus lactulose (n = 31) and lactulose alone (n = 31) in MHE patients by testing neurocognitive function (NCT-A and DST), blood ammonia, liver function (ALT, AST and TBIL) and blood inflammation (IL-1β, IL-6 and TNF-α). Afterward, we detected NO, inflammatory cytokines (IL-1β, IL-6 and TNF-α) and the phosphorylation of P65, JNK, ERK as well as P38 in LPS-activated rat primary bone marrow-derived macrophages (BMDMs), peritoneal macrophages (PMs), and mouse primary BMDMs/PMs/microglia/astrocytes, to investigate the underlying mechanism of BBD inhibiting inflammation through TLR4 pathway. Also, the survival rate of mice, liver function (ALT, AST), blood inflammation (IL-1β, IL-6 and TNF-α), inflammatory cytokines (IL-1β, IL-6 and TNF-α) and histopathological changes in the liver, brain and lung were measured to assess the anti-inflammatory effect of BBD on neurocognitive function in endotoxin shock/endotoxemia mice.


RESULTS
BBD combined with lactulose significantly ameliorated neurocognitive function by decreasing NCT-A (p＜0.001) and increasing DST (p＜0.001); inhibited systemic inflammation by decreasing IL-1β (p＜0.001), IL-6(p＜0.001) and TNF-α (p＜0.001); reduced ammonia level (p = 0.005), and improved liver function by decreasing ALT(p = 0.043), AST(p = 0.003) and TBIL (p = 0.026) in MHE patients. Furthermore, BBD inhibited gene and protein expression of IL-1β, IL-6 and TNF-α as well as NO in rat primary BMDMs/PMs, and mouse primary BMDMs/PMs/microglia/astrocytes in a dose-dependent manner. BBD inhibited the activation of mouse primary BMDMs/PMs/microglia/astrocytes by regulating TLR4 pathway involving the phosphorylation of P65, JNK, ERK and P38. Also, BBD reduced the mortality of mice with endotoxin shock/endotoxemia; serum levels of ALT, AST, IL-1β, IL-6 and TNF-α; gene expression of IL-1β, IL-6 and TNF-α in the liver, brain and lung, and tissue damage in the liver and lung.


CONCLUSION
Our study provided for the first time clinical and experimental evidence supporting the use of BBD in MHE, and revealed that BBD could play a crucial role in targeting and regulating TLR4 inflammatory pathway to improve neurocognitive function in MHE patients.
['Lu|Bingjie|B|', 'Wu|Chao|C|', 'Azami|Nisma Lena Bahaji|NLB|', 'Xie|Dong|D|', 'Zhao|Changqing|C|', 'Xu|Wan|W|', 'Hui|Dengcheng|D|', 'Chen|Xi|X|', 'Sun|Runfei|R|', 'Song|Jingru|J|', 'An|Yongtong|Y|', 'Li|Kun|K|', 'Wang|Huijun|H|', 'Ye|Guan|G|', 'Sun|Mingyu|M|']
D000368:Aged D000818:Animals D000893:Anti-Inflammatory Agents D001253:Astrocytes D001921:Brain D002478:Cells, Cultured D002681:China D003071:Cognition D016207:Cytokines D004195:Disease Models, Animal D004365:Drugs, Chinese Herbal D019446:Endotoxemia D005260:Female D006501:Hepatic Encephalopathy D006801:Humans D018836:Inflammation Mediators D008264:Macrophages D008297:Male D008810:Mice, Inbred C57BL D017628:Microglia D008875:Middle Aged D011247:Pregnancy D013997:Time Factors D051197:Toll-Like Receptor 4 D016896:Treatment Outcome
Neurocognitive function MHE BBD TLR4 pathway Systemic inflammation Acrylamide (PubChem CID: 6579) Ammonium persulfate (PubChem CID: 62648) Carbon tetrachloride (PubChem CID: 5943) Dimethyl sulfoxide (DMSO) (PubChem CID: 679) Disodium hydrogen phosphate (PubChem CID:24203) Ethanol (PubChem CID: 702) chloroform (PubChem CID: 6212) Formaldehyde (PubChem CID: 712) Glycine (PubChem CID: 750) Methanol (PubChem CID: 887) Neuroinflammation Potassium chloride (PubChem CID: 4873) Potassium dihydrogen phosphate (PubChem CID: 516951) Sodium chloride (PubChem CID: 5234) Sodium dihydrogen phosphate (PubChem CID:23672064) Sodium dodecyl sulfate (PubChem CID: 3423265) Thioacetamide (PubChem CID: 2723949) Tris (PubChem CID: 6503) Xylene (PubChem CID: 6850715)