Based On Gut-liver Axis Study The Mechanism Of Chlorogenic Acid And Geniposide On Nonalcoholic Steatohepatitis

Objective:To observe the effect of chlorogenic acid and geniposide on rats with non-alcoholic steatohepatitis(NASH)rats,and to explore the mechanism of action of G on NASH based on the gut-liver axis.Methods:Thirty male Sprague-Dawley rats were divided into the normal group,the model group,the chlorogenmc acid and geniposide group,the pioglitazone hydrochloride group and the bifico group according to the random number table method.Rats in the model group,the chlorogenic acid and geniposide group,the pioglitazone hydrochloride group and the bifico group were given high-fat diet for 16 weeks to establish NASH model.The corresponding drugs were given at the beginning of 9th week,for a total of 8 weeks.After treatment,the level of triglyceride(TG),total cholesterol(T-CHO),AST activity in plasma was detected by biochemical method.The pathological changes of liver and intestinal tissue were observed by HE staining.The expression levels of inflammatory factors such as interleukin-1?(IL-1?),interleukim-6(IL-6)and tumor necrosis factor-a(TNF-a)in liver tissues,intestinal secretory immunofglobulin(sIgA)in Intestinal tissue and plasma D-lactic acid were detected by ELISA.End point chromogenic assay was used to detect the lhpopolysaccharide(LPS)levels in plasma.Changes in gut microbiota composition were observed with DNA amplification and high-throughput sequencing.Expression levels of toll-like receptor 4(TLR4)-nuclear transcription factor(NF)-?B p65 signaling pathway related proteins(TLR4,phosphorylated NF-KBp65?nuclear transcription factor-activated protein-1(ap-1)and myeloid differentiation factor 88(MyD88)were detected by Western blotting.Results:Compared with the normal group,the liver histopathological sections of the model group showed steatosis,liver cell swelling,hepatocyte ballooning,and diffuse inflammatory cell infiltration in the lobules.Which showed typical histologic features of NASH.The NAS integral>5,which indicated the estabish of NASH rat model.Compared with the model group,the steatosis,hepatocyte ballooning,and inflammatory infiltration were significantly reduced in the chlorofgenic acid-gardenoside group.the steatosis,hepatocyte ballooning,and inflammatory in the rats of pioglitazone hydrochloride and bifico group were improved compared with the model group.Compared with the normal group,the ileal epithelial cells of the model group were slightly exfoliated and the intestinal mucosal barrier was damaged.After the treatment with chlorogenic acid and gardenoside,the intestinal mucosal epithelial cells of rats were not exfoliated,and the structure of ileum mucosa was more complete than that of the model group.Compared with the normal group,the levels of fat weight,liver wet weight,liver index,liver TG content,LPS in plasma,sIgA in Intestinal tissue,plasma D-lactic acid,the activity of AST and the levels of inflammatory factors such as IL-1?,IL-6,TNF-a were significantly increased in the model group.After treatment,the levels of fat weight,liver wet weight,liver index,liver TG content,LPS in plasma,sIgA in Intestinal tissue,plasma D-lactic acid,the activity of AST and the levels of inflammatory factors such as IL-1?,IL-6,TNF-a were decreased respectively.In the model group,the abundance of Proteobacteria was significantly increased compared with the normal group.After treatment,the abundance of Proteobacteria in the chlorogenic acid-germanoid group was decreased.The expression of TLR4,AP-1,MyD88 and phosphorylated NF-?Bp65 in the liver of the model group was significantly increased.After treatment,the liver of the chlorogenic acid-germanoid group was TLR4,AP-1.The expression of MyD88 and phosphorylated NF-?Bp65 was significantly reduced.Conclusion:The study found that Geniposide could significantly improve the intestinal mucosal structure,reduce the abundance of Proteobacteria to correct the disturbances of microbiota to reduce endogenous endotoxin production,thereby significantly reducing plasma LPS levels.The expression of IL-1?,IL-6 and TNF-a in liver tissues of NASH rats was significantly decreased by LPS-TLR4 signaling pathway.It is suggested that the mechanism of action of Geniposide in the treatment of NASH is related to the regulation of gut-liver axis,improvement of intestinal mucosal barrier,reducing the production endogenous LPS,and reducing the expressionof inflammatory factor.