The Effects Of CTGF On The Expressions Of MMP-2, MMP-9, TIMP-1 And TIMP-2mRNA And The Concentration Of Collagen-IV In Rat Glomerular Mesangial Cells

Objective:(1)To investigate the effects of different glucose concentration on the expression of connective tissue growth factor(CTGF) and type IV collagen(Col-IV) in rat mesangial cells(RMCs).(2)To investigate the effects of CTGF on the expression of matrix metalloproteinase-2(MMP-2),MMP-9, tissue inhibitor of metalloproteinase -1(TIMP-1), and TIMP-2 in RMCs.(3)To study the effects of different concentration of CTGF on the secretion of Col-IV in RMCs.(4)To explore the influence of CTGF on the expression of MMPs and TIMPs,and its possible mechanisms in diabetic nephropathy.Methods:RMCs were subject to different concentration of glucose respectively,Semi-quantitative RT-PCR analysis was used to determined the RNA expression level of CTGF.RMCs were subject to different concentration of CTGF respectively, Semi-quantitative RT-PCR analysis was used to determined the RNA expression level of MMP-2 ,MMP-9 ,TIMP-1 and TIMP-2, enzyme linked immunosorbnent assay(ELISA) method was used to determined the concentration of Col-IV.Results(1)There was no significant difference between the expression of CTGFmRNA at 5.6 and 11.0mmol/L of glucose concentration with the increase of time. The expression of CTGFmRNA was upregulated by 25mmol/ L concentration of glucose in a time-dependent manner.(2)The expression of TIMP-1mRNA was upregulated by CTGF in a dose-dependent manner ( p<0.05 ) and the expression of MMP-2mRNA and MMP-9mRNA were down-regulated by CTGF in a dose-dependent manner. CTGF has no influence on the expression of TIMP-2mRNA .(3)Exogenous CTGF could increase the concetration of Col-IV in a dose-dependent manner . CTGF could increase the concentration of Col- IV significantly at the level of 2.5ng/ml(P<0.05).Conclusions(1)The expression of CTGFmRNA could be upgregulated by 25mmol/ L concentration of glucose in a time-dependent manner.(2)The expression of MMP-2mRNA and MMP-9mRNA were down-regulated by CTGF in a dose-dependent manner , and TIMP-1mRNA was upregulated by CTGF in a dose-dependent manner.(3) There was a negative correlation between Col-IV and the epression of MMP-2mRNA and MMP-9mRNA,while there was a positive correlation between TIMP-1 and Col-IV.CTGF could increase the concentration of Col-IV and play an important part in diabetic nephropathy.