Effects of rosiglitazone on the expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in renal interstitial fibroblasts induced by cyclosporine A / 中华肾脏病杂志

ABSTRACT

Objective To investigate the effects of rosiglitazone on the expression of peroxisome proliferator-activated receptor-γ (PPARγ) and matrix metalloproteinase-9 (MMP-9) in renal interstitial fibroblasts induced by cyclosporine A, and discuss renal protective effect of rosiglitazone on renal toxicity of cyclosporine A. Methods Construction, screening and amplification of the target siRNA vector for PPARγ were carried out.The inhibitory effect of siRNA on the expression of PPARγ in normal rat kidney (NRK) cells was evaluated.NRK cells were cultured in the routine way.Experimental groups: (1)control group:single NRK cells without treatment; (2)RGZ group:NRK cells with RGZ (10 μmol/L); (3)CsA group:NRK cells with CsA (1.0 mg/L); (4)CsA+RGZ group:NRK cells with CsA (1.0 mg/L) plus RGZ (10 μ mol/L); (5)CsA+RGZ+siRNA group:pRNAT-U6.2/Lenti-PPARγ-236 plasmid transfected into NRK cells,then CsA (1.0 mg/L) plus RGZ (10 μmol/L).The mRNA expression of PPARγ was detected by real-time RCR.The mRNA expressions of MMP-9 and TIMP-1 were detected by RT-RCR.The protein expression of FN was detected by Western blotting. Results CsA up-regulated the mRNA level of PPARγ,MMP-9 and TIMP-1 (P＜0.05),and the up-regulation was inhibited by RGZ significantly (P＜0.05).The application of PPARγ siRNA resulted in the decreasing of PPARγ mRNA (P＜0.05),and partly reversed the inhibition effect of RGZ on MMP-9 and TIMP-1 mRNA (all P＜0.05).CsA up-regulated the protein level of FN (P＜0.05),and this effect was significantly inhibited by RGZ (P＜0.05).The application of PPARγ siRNA could reverse the inhibition effect of RGZ on FN protein expression (P＜0.05). Conclusion RGZ can inhibit the expressions of FN,MMP-9 and TIMP-1 induced by CsA which may be the mechanism of the protective effect of RGZ on renal interstitial fibrosis induced by CsA.