Based On Intestinal Microecology To Study The Intervention Mechanism Of Xiaoyao San On Liver Depression And Spleen Deficiency Of Non-alcoholic Fatty Liver Disease

Objective : Based on a large number of clinical observation and pre-experimental research in the early stage,based on the intestinal microecology,this paper discusses the prevention and treatment effect of Xiaoyao San,which has the functions of harmonizing liver and spleen,relieving liver depression,nourishing blood and strengthening spleen,on the model rats of non-alcoholic fatty liver disease(NAFLD)with liver depression and spleen deficiency,and studies the mechanism of its treatment on NAFLD.Methods :(1)Model building:44 male rats were randomly divided into 3 groups,including 10 in blank control group,10 in NAFLD group and 24 in NAFLD liver depression and spleen deficiency group.The blank control group was fed with common feed(standard feed),and the other two groups were fed with high-fat emulsion.The rats in NAFLD liver depression and spleen deficiency group were restrained for 4 hours,and clipped tail for 1 hour every day,and their body weight was measured once a week,and the dose of gavage was 1.5ml/100 g.After the 8th week,a model group of rats was randomly killed and evaluate the success of modeling.(2)Drug treatment:Elimination of rat death during modeling,according to the needs of the experiment,group C was subdivided into C1 group,C2 group and C3 group.There were 9 in the blank group,8 in the NAFLD model group,6 in the NAFL liver depression and spleen deficiency model group,6 in the NAFLD liver depression and spleen deficiency model of traditional Chinese medicine group,and 6 in the NAFLD liver depression and spleen deficiency model of Western medicine group.The blank group and model group were given 0.9% sodium chloride solution,the western medicine group was given atorvastatin calcium tablet and the traditional Chinese medicine group was given Xiaoyao San.The body weight was measured once a week,and the dose of gavage was adjusted according to the change trend of body weight.The drug intervention lasted for 2 weeks.On the day of the last administration,fecal samples were collected from the intestines.The rats were fasted for 12 hours after the last administration.On the second day,all rats were killed,and blood samples and liver samples were collected.(3)Detection index:In serum,aspartate aminotransferase,alanine aminotransferase,alkaline phosphatase,γ-glutamyl transpeptidase,total bilirubin,cholesterol,triglyceride,Low Density Lipoprotein,bile acid,glucose should be detected.Histopathological examination of liver:Hematoxylin eosin staining was used to evaluate the fatty degree of liver tissue in rats.The levels of IL-1 β,IL-6 and TNF-α in serum were detected by ELISA.16 S r RNA gene sequencing was used to detect diversity and abundance of intestinal flora in feces.Results:(1)According to the results of liver HE staining,NAFLD model group was found to have fatty changes in liver;according to the changes of hair yellowing,frequent squinting,stool characteristics,body weight and diet,NAFLD model of liver depression and spleen deficiency was successfully reproduced.(2)According to the changes of hair moistening,squinting improving,defecation forming,diet and body weight increasing,it was found that the symptoms of liver depression and spleen deficiency syndrome in rats with NAFLD treated with Xiaoyao San were improved.(3)According to the results of liver function,blood lipid and blood glucose in rats,it was found that Xiaoyao San and atorvastatin calcium tablets had no significant improvement on these indexes,but the degree of steatosis of hepatocytes in Xiaoyao San group was relieved to some extent.(4)According to 16 S r RNA The results of gene sequencing showed that the abundance of Clostridium in Xiaoyao San group was higher than that in atorvastatin calcium group(P < 0.05);the abundance of Clostridium in Xiaoyao San group was higher than that in atorvastatin calcium group(P < 0.05);the abundance of muribaculaceae in Xiaoyao San group was lower than that in atorvastatin calcium group(P < 0.05).There was significant difference between the Atorvastatin Calcium Tablets group and the Muribaculaceae group(P=0.006<0.01).At the genus level,the abundance of faalibacrum was higher in the Xiaoyao San group than that in the atorvastatin calcium group(P<0.05)(5)The results of ELISA showed that the level of IL-1 β in group C2 was significantly lower than that in group C1(P < 0.05),but there was no significant difference between group C2 and group C3.Conclusion: On the basis of the successful construction of NAFLD rat model of liver depression and spleen deficiency,after two weeks of intervention with Xiaoyao San and atorvastatin calcium tablet,the level of serum inflammation and the degree of liver steatosis in the two groups were improved,especially in Xiaoyao San group.Combined with the analysis of intestinal flora of rats in each group,it was found that Xiaoyao San could increase the level of beneficial bacteria and reduce the level of pathogenic bacteria,and there was a significant difference in species diversity and abundance between Xiaoyao San and atorvastatin calcium group,suggesting that Xiaoyao San may play a therapeutic role by regulating the intestinal flora of NAFLD rats.