Mechanism Of Chaihu Shugan Powder In The Treatment Of Cholesterol Gallstone Induced By High Fat Diet

Objective: To observe and analyze the relationship between high-fat diet and cholesterol gallstone,and to study the mechanism of Chaihu Shugan powder in intervening cholesterol gallstone caused by high-fat diet.Methods: The experiment was divided into five parts.The first part is an epidemiological investigation.The relationship between hyperlipidemia and incidence rate of cholecystolithiasis in 57977 physical examination centers of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine was analyzed.The second part is the experimental study of Chaihu Shugan powder in preventing cholesterol gallstone formation in mice fed with high-fat diet.30 mice were divided into blank group(fed with basic diet for 10 weeks),model group(fed with high-fat diet for 10 weeks)and treatment group(fed with high-fat diet for 10 weeks and intervened with traditional Chinese Medicine);Histological examination of bile,liver and gallbladder;The blood lipid and biochemical level of bile and serum were measured.The third part is the differential expression and screening of mi RNA related to the treatment of cholesterol gallstone in mice with Chaihu Shugan powder.In the blank group,model group and treatment group,the liver tissues of three mice were randomly selected for highthroughput sequencing of mi RNA,and the differential mi RNA,related target genes and signal pathways were screened and verified.In the fourth part,we verified the expression of differential mi RNA and related target genes in hepatocytes.Nine mice were divided into blank group,model group and treatment group,with 3 mice in each group.After 10 weeks of modeling,the liver tissues of mice were taken to isolate hepatocytes,and the expression levels of differential mi RNA and related target genes in hepatocytes were analyzed.In the fifth part,through the preparation of Chaihu Shugan powder containing serum lyophilized powder,we intervened AML12 cells and Hep G2 cells established by OA,observed the oil red O staining,TBA level,TC level,and the expression of mi R-871-3p and related target genes.At the same time,by regulating the expression of mi R-871-3p,we observed the expression level of cell related target genes after the intervention of medicated serum.Results: Hyperlipidemia was a high risk factor of gallstone,and high TC was an independent high risk factor of gallstone(P<0.001).Chaihu Shugan powder can effectively reduce the stone formation rate of cholesterol stones in mice induced by highfat diet(P<0.001),improve the liver tissue fat infiltration and gallbladder wall inflammation and edema,regulate liver cell metabolism,improve blood lipid and liver function levels(P<0.05).High throughput sequencing showed that the therapeutic effect of Chaihu Shugan powder on cholesterol gallstone caused by high-fat diet may be related to the gene regulation function of mir-871-3p and mir-465a-3p.The corresponding target genes of mir-871-3p were Akr1d1、Cyp27a1、Ldlr、Hmgcr、Slc10a2 and Npc2;the target genes of mir-465a-3p are Ldlr、Hmgcr、Slc10a2 and Npc2.These mi RNAs and related target genes are expressed in hepatocytes.After intervention with Chaihu Shugan powder,the expression of mir-871-3p was up-regulated in AML12 cells and Hep G2 cells,and the expression of corresponding key target genes Akr1d1 and Cyp27a1 was down regulated(P<0.05),which was consistent with the regulatory effect of Micic on enhancing mi RNA,and contrary to the regulatory effect of Inhibitor on inhibiting mi RNA.Conclusion: 1.Hyperlipidemia is a high risk factor of cholecystolithiasis;High TC is an independent risk factor for gallstone.2.Chaihu Shugan powder can effectively reduce the stone formation rate of cholesterol stones in mice induced by high-fat diet,protect hepatocyte degeneration,reduce fat vacuoles,improve gallbladder wall inflammation and edema,regulate hepatocyte metabolism,improve blood lipid and liver function.3.The therapeutic effect of Chaihu Shugan powder on cholesterol stones caused by high-fat diet may be related to the gene regulation function of mir-871-3p and mir-465a-3p.Akr1d1,Cyp27a1、Ldlr、Hmgcr、Slc10a2 and Npc2 are potential LITH genes.4.Chaihu Shugan powder can effectively improve the lipid infiltration and bile acid metabolism of AML12 cells and Hep G2 cells,and its mechanism of preventing cholesterol stones may be related to up regulation of mi R-871-3p and down regulation of Akr1d1 and Cyp27a1 expression levels.