| Objective: To detected the expression of miR-196 a and its potential target gene ANXA1?PTEN in diabetic nephropathy(DN)cells and mouse kidney tissue,explored the relationship between them with diabetic nephropathy.Methods:(1)The mouse glomerular mesangial cells was treated with high glucose,replicated the cell model of diabetic nephropathy,detected the gene expression of ?-smooth muscle actin(?-SMA)and fibronectin(FN)by qRT-PCR,Western blot and ELISA.(2)The potential target gene of miR-196 a was predicted by bioinformatics software,and its mRNA expression was detected by qRT-PCR.(3)miR-196 a inhibitor was transfected into diabetic nephropathy cells,the expression of ANXA1 and PTEN was detected by qRT-PCR and Western Blot.The cell proliferation was detected by CCK-8 assay.(4)Copied animal model of diabetic nephropathy,detected the expression of candidate miRNA in renal tissue of DN mice.The expression of ANXA1 and PTEN was detected by qRT-PCR and Western blot.Results:(1)When glucose concentration was 75mmol/L and intervented time was 48 h,the expression of ?-SMA and FN was significantly higher than control group,and the difference was statistically significant(P<0.05),prompted the cell model of diabetic nephropathy was successfully reproduced.(2)Compared with the NC group,the expression of miR-196a?miR-98 was significantly higher in DN group,and the difference was statistically significant(P<0.05).(3)Compared with the NC group,the expression of ANXA1 and PTEN was significantly lower in DN group,and the difference was statistically significant(P<0.05).miR-196 a inhibitor was transfected into DN cells,the expression of ANXA1 and PTEN compared with the DN group was significantly increased(P<0.05).(4)CCK-8 assay showed the OD value of mesangial cells in DN group was higher than NC group(P<0.05),the OD value of mesangial cells in miR-196 a inhibitor group was significantly lower than DN group,and the difference was statistically significant(P<0.05).(5)Compared with the blood glucose and 24 h urinary protein excretion rate in model group and control group,prompted the animal model of diabetic nephropathy was successfully reproduced.(6)The expression of miR-196a?miR-21 was significantly increased in DN mice compared with NC mice,and the difference have statistically significant(P<0.05).The expression of miR-196 a was positively correlated with the level of blood glucose,triglyceride,total cholesterol and 24 h urinary protein excretion rate in DN mice(P<0.05).(7)The protein expression of ANXA1 and PTEN in kidney tissue of DN group was significantly lower in NC group,and the difference was statistically significant(P<0.05).Conclusion: miR-196 a may involve in the development of diabetic nephropathy by regulating the expression of ANXA1 and PTEN. |
PDF Full Text Request