Preliminary Study On The Regulator Role Of MicroRNA-98 For SOCS1 And ZEB2 In The Diabetic Nephropathy

Objective:To detect the expression level of microRNA-98(miRNA-98)and it's target gene SOCS1?ZEB2 in the diabetic nephropathy(DN)cell and mouse model,explore the regulator role of miRNA-98 for it's target gene in the DN.Methods:(1)The DN cell replication model was constructed by culture mouse mesangial cells in the high glouse condition,the expression levels of 7 candidate miRNA was detected by quantitative Real time PCR(qRT-PCR),among them miRNA-98 was chosen to study.(2)Predict the target gene of miRNA-98 by bioinformatic analysis,SOCS1?ZE B2 mRNA and protein levels were detected by qRT-PCR?western Blot.(3)miRNA-98 mimics and inhibitor was transfected into DN cells,SOCS1?ZEB2 mRNA and protein levels were detected by qRT-PCR?western Blot.(4)Replicate the DN mouse model,the expression levels of 7 candidate miRNA was detected by qRT-PCR,mRN A and protein levels of miRNA-98 target genes SOCS1?ZEB2 was detected by qRT-PCR?western blot.Results:(1)Compared with control group,the expression levels of miRNA-98?miRNA-196 a was increased highily in DN cell group(P?0.05),while miRNA-709?miRN-21?miRNA-187?miRNA-451?miRNA-451 were not significantly different(P?0.05).(2)After target peredicrion of miRNA-98 by using miRDB?microRNA.org,there were 430?5031 target genes were collected,SOCS1 and ZEB2 which were closely associated with DN were selected from 363 comon the targrt genes and mmu-miR-98 has one complementary binding site in the 3'-UTR of SOCS1?ZEB2 respectively.(3)The mRNA and protein levels of SOCS1?ZE B2 in the DN group were lower than in the control group(P?0.05).The mRNA and protein levels of SOCS1?ZEB2 in the miRNA-98 mimics group were lower than in the DN group(P?0.05),while higher in the miRNA-98 inhibitor group(P?0.05).(4)At week 8,db/db mouse showed the heperglycemia and significant elevation of 24 h urinary albumin exceretion rate compared with db/m mouse,proved that DN mouse model were replicated successfully.Compared with control group,miRNA-196a?miRNA-21 were highly expressed in the renal tissue of DN mouse(P?0.05),while miRNA-98?miRNA-187?miRNA-709?miRNA-451?miRNA-29 cexpression levels were not significantly different(P?0.05)?The mRNA and protein levels of SOCS1?ZEB2 were reduced in DN group campared with control group(P?0.05).Conclusion:miRNA-98 expression was upregulated in the diabetic nephropathy cell model,the results of cell and mouse model experiment suggest that SOCS1 and ZEB2 were targeted by miRNA-98,might be promote extracellular matrix accumulation and fibrosis of mouse mesangial cells,caused the pathogenesis of diabetic nephropathy.