Study Of Targeting CD248 In Interfering With Hepatic Fibrosis In Mice

BackgroundHepatic fibrosis is characterized by activation and proliferation of hepatic stellate cells(HSC)and excessive deposition of extracellular matrix(ECM),which is the final outcome of many chronic liver diseases.It is of great significance to explore non-invasive biomarkers of liver fibrosis and effective intervention strategies.The expression of CD248 can be detected in fibrosis.Knocking out the CD248 gene can inhibit the process of fibrosis,indicating that CD248 is related to the development of fibrosis and other diseases,but the specific mechanism of action of CD248 in fibrosis is still unclear.Objectives1.Through the pre-made CD248 monoclonal antibody,explore the possibility of CD248 molecule as a biomarker of liver fibrosis;2.Explore whether antibody neutralization interferes with CD248 molecule to affect the development of fibrosis,and then determine whether CD248 can be used as a target for fibrosis diagnosis and treatment.Methods1.Detection of CD248 expression:collect serum from normal medical examinees and patients with liver cirrhosis,and pathological specimens from patients with cirrhosis;construct a mouse liver fibrosis model and use the self-made anti-CD248 monoclonal antibody of the research team to detect the expression of CD248.2.Targeting CD248 to interfere with hepatic fibrosis in mice:Mice were randomly divided into three groups,and the mice in the antibody group and the model group were injected intraperitoneally with 25%carbon tetrachloride(dissolved in olive oil)to induce liver fibrosis.The normal group accepted the same dose of olive oil.From the second week of injection,the anti-CD248 monoclonal antibody was injected intravenously into the antibody group twice a week for a total of 6 times.Mice were sacrificed at the end of the 6th week of carbon tetrachloride injection,and serum and liver were collected.AST and ALT levels in serum of mice were detected,HE and Sirius red staining were used to observe the degree of hepatic fibrosis,Western Blot,immunohistochemistry and qRT-PCR were used to detect ?-SMA and Collagen I protein and mRNA levels in liver.3.Preliminary study on the mechanism of CD248 participating in liver fibrosis:Western Blot,immunohistochemistry and qRT-PCR were used to detect VEGF and HIF-1? protein and mRN A levels in the liver.Results1.Western blot results showed that CD248 level in serum of cirrhosis patients was higher compared to normal people,but there were individual differences.Immunohistochemical results showed that CD248 was highly expressed in liver cirrhosis tissues.High expression was detected in the mouse liver fibrosis model.2.Liver function tests and HE and Sirius Red staining results showed that the levels of AST and ALT in the model group were significantly increased compared to the normal group,with significant fibrosis(P<0.05);the AST and ALT levels in the antibody group was decreased compared to the model group,and the degree of fibrosis was reduced(P<0.05).Western Blot,immunohistochemistry and qRT-PCR results showed that the expression levels of ?-SMA and Collagen I in the liver of the antibody group were decreased compared to the model group(P<0.05).After the mice in the antibody group were given antibodies,the expression of CD248 decreased.3.Western Blot,immunohistochemistry and qRT-PCR results showed that the expression levels of VEGF and HIF-1? in the liver of the antibody group were reduced compared to the model group(P<0.05).Conclusions1.Using self-made antibodies can detect the increase of CD248 serum in patients with liver cirrhosis and the increase in mouse liver fibrosis model.CD248 molecule can be used as a biomarker for liver fibrosis.2.The intervention of anti-CD248 antibody to reduce the formation of liver fibrosis in mice may be related to the influence of HSC activation and proliferation and the reduction of vascular proliferation.CD248 molecule may become a new target for the treatment of liver fibrosis.