Study On The Relationship Between Intestinal Flora,Beta 2-glycoprotein ? And Hepatic Fibrosis

Objective:Recently,the cirrhosis has been becoming the most common cause of death in the world,as the incidence and mortality of cirrhosis is rising.Liver fibrosis is an inevitable pathological process for the development of chronic liver disease into cirrhosis,because the liver fibrosis is a reversible process,so intervention and treatment as soon as possible are essential for the development of liver disease.Currently,there are no specific drugs for the treatment of liver fibrosis,therefore,taking the etiology of liver fibrosis as the starting point to understand the mechanism of liver fibrosis development is of great significance for the treatment and prevention of liver fibrosis.The purpose of our study was to investigate the relationship between intestinal flora,beta2-glycoprotein I(beta2-GP?)and liver fibrosis.We first studied the changes of intestinal flora in mice with liver fibrosis,and then we analyzed the levels of ?2-GP?,Nuclear factor-KB p65(NF-?B p65)and some coagulation indexes such as activated partial thromboplastin time(APTT),prothrombin time(PT)and the activity of coagulation factor XI(FXIa),followed the relationship between gut flora,?2-GP? and liver fibrosis was discussed.Meanwhile,the serum levels of ?2-GP? and plasma APTT,PT and the FXIa from patients with cirrhosis and healthy people were analyzed to discuss the relationship between ?2-GP? and liver fibrosis.Methods:1.Animal experiments:(1)Establish the liver fibrosis models:The sixteen BALB/c mice were randomly divided into two groups(control group and liver fibrosis group),the mice were received CCL4 and peanut oil mixture by intraperitoneal injection to establish the liver fibrosis models.(2)Samples collection:The mice were sacrificed at 8 weeks post-infection of CCL4,and samples of liver,blood and cecal contents were collected.(3)Detect and analyze samples:We analyzed the serum levels of aminotransferase and ?2-GP? in mice,and the APTT,PT,FXIa in mice plasma were detected;besides,we also observed the pathological changes and the expression of ?2-GP? and NF-?B p65 of liver tissue;in addition,we analysed the gut microbiota diversity using I-Sanger biodata analysis platform.2.Clinical experiments:The blood samples from fifty patients with cirrhosis and fifty normal healthy people were collected and the serum levels of ?2-GP? and plasma APTT,PT and the FXIa were tested.3.Statistical method:Analysis with SAS 9.2 statistical software.Measurement data were mean ± standard(x±s)deviation for statistical description,T test was adopted to compare measurement data between two groups.And P<0.05 was considered statistically significant.Results:1.Animal experiments:(1)Compared with control group,the serum of alanine aminotransferase(ALT),aspartate aminotransferase(AST)of the hepatic fibrosis group were declined obviously,while there was no significant difference about the gamma-glutamyltransferase(y-GGT)between the control group and the hepatic fibrosis group.Histochemical staining of liver tissues showed that hepatic lobule destruction,hepatocyte apoptosis and inflammatory cell proliferation and collagen deposition were significantly increased in hepatic fibrosis group,these results indicated that the model of hepatic fibrosis have been successfully established.(2)Alpha diversity of intestinal community decreased in hepatic fibrosis group,and intestinal flora imbalance characterized by the sharp increased abundance of the intestinal bacteria such as Actinobacteria and Desulfovibrionaceae and the like,and a significant decrease in the abundance of the Lachnospiraceae.(P<0.05)(3)The serum level of[32-GP? in hepatic fibrosis group was obviously lower than that in normal control group(P<0.05),while there was no significant difference in the expression of ?2-GP? in liver tissue between control group and hepatic fibrosis group.Also,compared with control group,the expression of phosphorylated NF-?B p65(P-NF-?B p65)in liver tissue were markedly increased(P<0.05),and the expression of total NF-?B p65(NF-?B p65)in hepatic fibrosis group liver tissues were comparable with that in control group.In liver fibrosis group the FXIa decreased dramatically when compared with the control group,but APTT and PT have no significant difference between the two groups.2.Clinical experiments:The serum ?2-GP? and FXIa declined obviously in cirrhosis patients,and compared with normal healthy people there was no significant difference in APTT of cirrhosis patients,but the PT greatly elevated in cirrhosis patients.Conclusion:1.Alpha diversity of intestinal flora decreased,intestinal permeability increased,intestinal flora imbalance in liver fibrosis mice.2.The imbalance of intestinal flora may induce the downregulation of ?2-GP? through microbial associated molecular patterns(MAMPs),and then initiated the NF-?B p65 and ultimately leaded to the development of liver fibrosis.Meanwhile,we found that the FXIa declined significantly,but did not affect APTT,providing new ideas for clinical thrombosis treatment.