Protective effect and mechanism of Fufang Zhige Yin on chronic alcoholic liver injury based on NF-κB/NLRP3 pathway and metabolomics
Objective:To explore the therapeutic effect of Fufang Zhige Yin on alcoholic liver injury and its mechanism based on NF-κB/NLRP3 pathway and metabolomics.Methods:The mice were divided into blank group,model group,group,high and low dose of Fufang Zhige Yin water extract,and high and low dose of Fufang Zhige Yin alcohol extract.In addition to the blank group,the rest of the groups were given alcohol gavage to prepare the alcoholic liver injury model,and at the same time,the corresponding drug treatment was given.After 8 weeks,the liver ALT,AST,TC,TG,SOD,MDA indexes were detected in mice,HE staining was used to observe the pathological changes in the liver of the mice,and the serum levels of inflammatory factors TNF-α,IL-1 β,and IL-18 were detected in the mouse serum by ELISA method,and the levels of the inflammatory factors TNF-α,IL-1 β,and IL-18 were detected in the liver of the mice by qRT-PCR to detect mRNA expression of TNF-α,IL-6,IL-1 β genes in mouse liver,Western Blot was used to detect NF-κB and NLRP3 protein expression in liver,and LC-MS non-targeted metabolomics to analyze the changes of endogenous metabolites and pathways in serum and liver.Results:Compared with the model group,both the extracts of the Fufang Zhige Yin water and the alcoholic extracts significantly reduced the hepatic ALT,AST,TC,TG,serum TNF-α levels,hepatic expression of inflammatory gene IL-1 β mRNA levels,hepatic expression of NF-κB protein and NLRP3 protein in mice(P＜0.05,P＜0.01);the aqueous extracts also significantly elevated the SOD activity of mice and significantly reduced the mouse liver MDA and TNF-α mRNA expression(P＜0.05,P＜0.01);the alcoholic extract also significantly reduced serum IL-18 levels(P＜0.05);serum metabolomics screened for 10 metabolites associated with alcoholic extract for the treatment of alcoholic liver injury,of which 6 could be up-regulated and 4 could be down-regulated,and 12 aqueous extracts for the treatment of alcoholic liver injury metabolites,of which 6 could be up-regulated and 6 down-regulated;liver metabolomics screened out 21 liver metabolites related to aqueous extracts for the treatment of alcoholic liver injury,of which 13 could be up-regulated and 8 down-regulated,and the metabolic pathways involved were mainly the following:ascorbic and aldic acid metabolism,purine metabolism,arginine biosynthesis and other pathways,and screened out 21 liver metabolites related to alcoholic extracts for the treatment of alcoholic liver injury,of which 2 could be up-regulated and 19 down-regulated,and the metabolic pathways involved were mainly:starch and sucrose metabolism,taurine and low taurine metabolism,ascorbate and aldolate metabolism,etc.There were 8 metabolites that could be regulated by both aqueous and alcoholic extract;and the metabolic pathways that could be acted on by both aqueous and alcoholic extract were ascorbic and aldolacic metabolism,sphingolipid metabolism,starch and sucrose metabolism.Conclusion:Both aqueous and alcoholic extracts of Fufang Zhige Yin have therapeutic effects on the treatment of alcoholic liver injury,and their mechanism of action may be through the regulation of NF-κB/NLRP3 signaling pathway to reduce the level of inflammatory factors,as well as the regulation of serum and hepatic metabolite disorders.
NETL
NSTL
万方数据
