Study On The Network Of Efficacy And Mechanism Of Xiao Chai Hu Decoction In The Treatment Of Nonalcoholic Steatohepatitis

Objective:This study was designed to investigate the efficacy network and mechanism of Xiaochaihu Decoction in treatment of non-alcoholic steatohepatitis（NASH）,in order to provide experimental evidence for the NASH study of new treatment strategies and clinical precision medication of Xiaochaihu Decoction.Methods:In C57BL/6 mice for the study,administration of methionine choline deficient（MCD）diet induced mouse model of NASH.Observing the state of mice,studying liver tissue pathological changes by HE staining,analysing the changes of lipid metabolism and inflammation related indexes in serum and liver tissue,and exploring its lipid-lowering mechanism by q RT-PCR experiments.Free fatty acids（FFA）were induced fat cells L-02 model to observe the effect Xiaochaihu Decoction extract on lipid deposition and the extent of damage to cells.Using network pharmacology technology built a“disease-prescription–target”network,predicted Effective Compounds and the key target for the Xiaochaihu Decoction in treatment of NASH,and carried on Go and pathway enrichment analysis for the key targets to predict the potential efficacy network.last but not least,verifying the related indexes of Action Network by way of q RT-PCR experiments Results:Xiaochaihu Decoction can significantly improve hepatic steatosis and inflammation and other changes,reduce TC and TG levels in serum and liver tissue,reduce the secretion of MDA and SOD,reduce AST,ALT and LDH concentration in serum.q RT-PCR experiments results showed that Xiaochaihu Decoction could reduce the expression of FAS,SREBP-1c,SREBP-2,HMG-Co A and TNF-α,IL-6 and IL-1β,which were related to liver fat metabolism.Establishment of L-02 high fat cell model induced by 400mmol·L^-1FFA.0.625～2.50 mg·m L^-1Xiaochaihu Decoction extract can significantly improve the rate of fat cells steatosis,and reduce TC and TG levels in the cell supernatant levels of ALT,AST,TNF-αand IL-6 expression of inflammatory cytokines in cells.Network pharmacology predicts that Xiaochaihu Decoction can interfere with NASH mainly by acting on 38 key targets such as NFKB2 and NFKBIA through 182 chemical components in the formula,participating in multiple pathways such as interleukin signal,immune system and activation of NF-κB mediated by TRAF6.By detecting the expression of related genes in liver tissue of mice in NASH groups,Xiaochaihu Decoction can significantly reduce the transcription factor p65/Rel A,P50/NFKB1 and p52/NFKB2,which were related to NF-κB pathway.Conclusions:In this study,in vivo and in research methods combined with network technology to carry out the pharmacological mechanism of action of therapeutic effect Xiaochaihu Decoction,Xiaochaihu Decoction clear in vivo and in vitro showed a significant level of liver lipid-lowering effect and a good dose-effect relationship.Fat or free fatty acid-induced hepatic steatosis can cause injury and expression of proinflammatory cytokines TNF-αand IL-6.Under the action of proinflammatory cytokines,TNF-αand NF-κB interaction may constitute a network,by of FAS,SREBP-1c isogenic,inflammatory cytokines and NF-κB pathway genes such as P65,P50 and P52.The detection results suggest that Xiaochaihu Decoction may play a role in reducing blood lipid and protecting liver of NASH by regulating genes related to fat metabolism,interleukin signaling pathway,immune system and NF-κB pathway.