Experimental Study Of CD248 Antibody Intervention On Renal Fibrosis In Mice

BackgroundChronic kidney disease(CKD)has become a global public health concern,and its morbidity and mortality have increased year by year.Fibrosis is a common pathway for a variety of CKD progressing to ESRD,and it is important to slow or reverse the fibrosis process.Despite the enormous clinical need,there are still few strategies for effective intervention in fibrosis.CD248 is a type ? transmembrane glycoprotein with a molecular weight of 175 kDa.It was found that CD248 is involved in the development of fibrosis,but the specific mechanism remains unclear.PurposeThe study intends to use the renal fibrosis mouse model as the research object,examine the related-indicators of fibrosis,analyze the effects of interfering CD248 molecular pathway in fibrosis,and then explore the mechanism of CD248 in fibrosis.Method1.A folic acid-induced renal fibrosis mouse model was established.C57BL/6 mice were randomly divided into normal control group and folic acid(FA)group,which were intraperitoneally injected with PBS or folic acid at a dose of 250 mg/kg.Mice were sacrificed on the 3rd,14th and 28th day after folic acid injection.Blood and kidney tissues were collected to measure Serum creatinine and UREA levels.HE and MASSON staining were used to observe the renal pathological changes and immunohistochemical staining was used to examine the protein levels of ?-SMA,Collagen I,Fibronectin and CD248 in kidney tissues.2.The intervention experiment with CD248 monoclonal antibody:the mice were randomly divided into normal control group,FA model group with PBS injection,FA model group with low-dose CD248 antibody injection and high-dose CD248 antibody injection.Beginning on the 3rd day after intraperitoneal injection,the mice were infused with PBS,2.5 mg/kg antibody or 7.5 mg/kg antibody by tail vein.Mice were sacrificed on the 28th day to examine the levels of Serum creatinine and UREA.HE and MASSON staining were used to observe the degree of renal fibrosis.The protein and mRNA levels of ?-SMA,Collagen I,Fibronectin and CD248 were examined by Western Blot,qRT-PCR and immunohistochemical staining.Result:1.Compared with the normal control group,Serum creatinine and UREA levels were significantly increased in the FA3 day,FA14 day,and FA28 day groups(P<0.05).The degree of renal fibrosis gradually increased.The protein expressions of ?-SMA,Collagen I,Fibronectin and CD248 in kidney tissues were increased.2.The level of Serum creatinine was significantly lower in the low-dose CD248 antibody group and the high-dose CD248 antibody group than the FA model group with PBS injection(P<0.05),but the level of UREA was not significantly decreased(P>0.05).Compared with the normal control group,HE and MASSON staining results showed that renal fibrosis was more obvious in the FA model group with PBS injection.The degree of renal fibrosis was significantly improved in the low-dose CD248 antibody group and high-dose CD248 antibody group than the FA model group with PBS injection.The improvement in the low-dose group was more obvious than the high dose group.Western Blot,qRT-PCR and immunohistochemical staining results showed that the protein and mRNA expressions of ?-SMA,Collagen I,Fibronect:in and CD248 in the low-dose CD248 antibody group and high-dose CD248 antibody group decreased significantly than the FA model group with PBS injection(P<0.05).Conclusion:1.A folic acid-induced renal fibrosis mouse model was established successfully.2.CD248 antibody intervention could improve renal fibrosis and CD248 may become an important target for fibrosis diagnosis and therapy.