The Effect Of Shugan Lipi Recipe On The Intestinal Flora Of Rats With Non-alcoholic Steatohepatitis

Objective: In this study,we studied the pathogenesis of NASH "spleen deficiency" from intestinal microecology,expounded the relationship between the theory of "liver disease and spleen" and intestinal mucosal homeostasis reconstruction,and explored the effect of Shugan Lipi Recipe on intestinal flora of NASH rat model.METHODS: Sixty healthy and clean male Sprague-Dawley(SD)rats were selected.Adaptive feeding for one week,divided into two groups,one group randomly selected 8 rats as the normal group,given normal diet.The other 52 rats were fed methionine choline deficiency diet(MCD diet),and the MCD diet was administered at 10g/d/100 g.Eight weeks later,two rats were randomly selected from the normal diet group(N),and four rats from the methionine-choline deficiency diet group were randomly selected to evaluate the pathological model of rats,and 36 fecal samples were collected from the two groups.The remaining 48 rats were randomly divided into six groups: model low dose group(ML),model high dose group(MH),model low dose control group(MLD),model high dose control group(MHD),model non-traditional Chinese medicine group(M)and model non-traditional Chinese medicine control group(MD).。 ML group and MH group were given MCD diet while traditional Chinese medicine was given orally.The MLD and MHD control groups were given traditional Chinese medicine and normal diet at the same time.In group M,MCD was continued,and in group MD,normal diet was given.The low and high doses of traditional Chinese medicine were given 0.92 g/day and 3.68 g/day respectively.After 4 weeks of treatment with traditional Chinese medicine,fecal samples of rats were collected.The whole experiment lasted for 12 weeks,during which the rats in each group were fed the same amount of food.The average weight of rats in each group was measured weekly.Result 1.General comparison At 8 weeks,the hair glow of the normal group was smooth,no hair loss was observed,body weight continued to increase and stool granules were big black.In the MCD diet group,the hair was dull,sparse,and the body weight continued to decrease.The stools were small and yellow.At 12 weeks,MH group and MD group improved significantly in body weight,hair and activity.Compared with MD group,MHD group improved significantly in body weight,hair and activity.2.HE staining of liver tissue In the MCD diet group,steatosis and degeneration of hepatocyte tissues were observed at different degrees after 8 weeks of modeling.Inflammatory cell infiltration and local small necrosis were observed.At 12 weeks after model establishment,hepatic pathology in group M showed balloon-like degeneration of hepatocytes,blurred boundary of hepatic lobules,inflammatory infiltration of hepatocytes,necrosis of hepatocytes,unclear boundary and hepatic fibrosis.Liver slices in MHD group showed that hepatic fat deposition was significantly less than that in the 8th week after operation.The boundary of hepatic lobule structure was clear,and no obvious inflammatory tissue infiltration was observed.3.High-throughput pyrophosphatic acid sequencing of 16 S rRNA in fecal intestinal flora 3.1 Species Composition AnalysisAll samples were annotated to 7 taxonomic levels,and 13 taxa were mainly annotated at the gate level.Among them,the main taxonomic levels were thick wall bacilli,bacteroides,Actinobacillus and denaturing bacilli,and the intestinal structure of MHD group and N group tended to be similar at the gate level.There were also significant differences in intestinal flora between MH group and M group at gate level.At the generic level,there was no significant difference in abundance between N group and MHD group.3.2 Alpha diversity analysis of rat intestinal fecesThere was no significant difference in richness(chao1)and diversity(shannon)between MLD group and MHD group,ML group and MH group(P > 0.05),indicating that the dosage of traditional Chinese medicine had no significant effect on the richness and diversity of intestinal flora.The richness and diversity of intestinal flora in MHD group were significantly different from those in MD group,but there was no significant difference between MHD group and MD group(P < 0.05).There was no significant difference between MHD group and normal group.3.3 Bae Diversity Analysis of Rat Intestinal Feces SamplesThe comparison of faecal intestinal flora between group N and the 8th week of modeling showed that there was significant difference in the structure of intestinal flora between the two groups(P < 0.05).Compared with MH group,the distance between MLD group and MHD group was far,and the intestinal flora structure was significantly different(P < 0.05).There was significant difference in intestinal flora structure between MHD group and MD group(P < 0.05).There was no significant difference between MHD group and N group(P > 0.05).There were significant differences in the abundance and diversity of intestinal flora between MH group and M group(P < 0.05).4.conclusion 1.NASH rat model was successfully induced by methionine-choline deficiency diet(MCD diet),which changed the structure of intestinal flora and damaged intestinal mucosal barrier.2.Simple low-dose intervention of traditional Chinese medicine had an impact on the abundance and diversity of intestinal flora,but there was no statistical significance.High-dose Chinese herbal medicine intervention has a significant impact on the abundance and diversity of intestinal flora,which can restore the abundance of intestinal flora in rats to a certain extent,reduce the diversity of intestinal flora,and regulate intestinal homeostasis.