Study On The Effect And Mechanism Of Jiang-Zhi Granules For NAFLD Through Upregulation Of Irisin Signaling Pathway

Objective To study the effect of Jiang-Zhi Granules in the treatment of NAFLD rats induced by high-fat diet and its mechanism and target based on Irisin signaling pathway.Methods 1.Twenty-four male wistar rats were randomly divided into three groups,control group,model group and Jiang-Zhi Granules(JZKL)group.The control group was given normal feed,the model and JZKL group were fed with high-fat diet for 8 weeks.From the 5th week of HFD,intragastric administration of Jiang-Zhi Granules was simultaneously given to the rats in the JZKL group.At the end of the experiment,the body weight,liver weight,liver histological pathology HE staining and oil red O staining,serum transaminases,lipid in liver and blood,as well as hepatic expression levels of Irisin,AMPK?,ACC,HMGCR m RNA and protein were evaluated.2.Induce the mouse 3T3-L1 preadipocytes into mature adipocytes with dexamethasone,isobutylmethyxanthine and insulin,to evaluate the effect of recombinant protein Irisin on adipogenic differentiation of 3T3-L1 preadipocytes.The m RAN and protein expressions of brown fat marker molecules CD137,TMEM26,PRDM16,mitochondrial molecules UCP1,Otop1,adipogenesis related molecules AMPK?,ACC,LXR?,SREBP1 C,PPAR?,FAS and fat oxidation related molecules PPAR?,CPT1 A were detected.3.Hepatocyte steatosis was induced by free fatty acids with Hep G2 cells.The recombinant protein Irisin combined with AMPK? inhibitor Compound C was used in Hep G2 cells to study the role of Irisin in hepatocytes of steatosis.The expression of AMPK?,ACC,HMGCR m RNA and protein and the phosphorylation of proteins of AMPK? and ACC were evaluated.Results 1.NAFLD rat model was established by eight-week HFD feeding.HE staining and oil red O staining showed that the fat deposition appeared in liver of NAFLD rats,the levels of serum ALT,AST,?-GT,TC,and LDL-c were significantly increased.Jiang-Zhi Granules significantly improved hepatic fat deposition,reduced serum transaminases and blood lipids in NAFLD rat,the mechanism may be involve up-regulation the expression of Irisin to reduce the lipid and cholesterol through Irisin-AMPK?-ACC and Irisin-AMPK?-HMGCR signaling pathway.2.Irisin could inhibit the adipogenic differentiation of 3T3-L1 precursor adipocytes,up-regulate the expression of white fat browning-related molecules CD137,TMEM26,PRDM16 m RNA and mitochondrial molecules Otop1 m RNA and UCP1 protein;down-regulate the expression of adipogenesis related molecules LXR?,SREBP1 c,ACC,PPAR? m RNA and PPAR? protein,increase the phosphorylation of AMPK? and ACC protein;up-regulate the m RNA and protein expression of PPAR? and CPT1.3.Irisin could reduce the fat and cholesterol in Hep G2 cells of steatosis by regulating the expression of ACC and HMGCR m RNA and protein,which is mediated by AMPK?.When Irisin and the AMPK? inhibitor Cmpound C were added to the hepatocellular steatosis at the same time,the effect of Irisin on ACC and HMGC disappeared.Conclusion 1.Jiang-Zhi Granules could up-regulate the expression of Irisin in the liver of NAFLD rats,reduce the lipid and cholesterol through Irisin-AMPK?-ACC and Irisin-AMPK?-HMGCR signaling pathway.2.Irisin could promote the transformation of mature 3T3-L1 adipocytes into brown adipocytes,regulate adipocyte energy metabolism through reduce lipid production and increase lipid oxidation.3.Irisin could reduce lipid and cholesterol in steatosis Hep G2 cells by regulating AMPK?,the mechanism may involve the Irisin-AMPK?-ACC and Irisin-AMPK?-HMGCR signaling pathway.