辛伐他汀对大鼠肾间质成纤维细胞的影响

EFFECTS OF SIMVASTATIN ON CULTURED RAT RENAL INTERSTITIAL FIBROBLASTS

目的 :研究 3 羟基 3 甲基戊二酰辅酶A(HMG CoA)还原酶抑制剂 (辛伐他汀 )对大鼠肾间质成纤维细胞 (RFB)增生 ,细胞周期及细胞间信号传导的影响。 方法 :应用MTT比色法、流式细胞仪、半定量RT PCR等技术 ,分别检测不同浓度辛伐他汀对体外培养的RFB的增生、细胞周期以及细胞信号传导中c fosmRNA表达的影响。 结果 :辛伐他汀可抑制RFB的增生活性 (A值 ) ,尤以无血清组显著 ,各浓度加药组A值与正常对照组比较差异显著 (P <0 0 5 ) ;各浓度加药组G0 /G1期细胞比增高 ,S期细胞比、PI值和DNA含量则逐渐下降 ,G2 /M期细胞比变化不明显 ;各浓度加药组RFBc fosmRNA的表达逐渐下降 ,并呈一定的剂量依赖性 ,与正常对照组比较差异显著 (P<0 0 5 )。 结论 :辛伐他汀可抑制细胞增生 ,阻止RFB由G1期进入S期 ,及阻断与细胞增生有关的c fos依赖性有丝分裂途径。辛伐他汀可能通过影响c fos依赖性有丝分裂传导途径抑制细胞增生。

Experi mental and clinical studies have demonst rated beneficial effects of 3-hydromxy-3 -methylgutaryl coenzyme A reductase inhi bitor on the structural and functional aspects of renal diseases.The present study is to investigate the effects of S imvastatin on cultured interstitial fibr oblasts from rat kidney(RFB),and the rel ated mechanism. METHODOLOGY Interstitial fibroblasts wer e separated from the medulla of rat kidn eys,and cultured and identified accordin g to the reported methods.Effects of Simv astatin on cell proliferation,cell-cycle and intracelluar signal transduction of RFB in primary culture,were measured by MTT assay,flow cytometry and semi-quant ity RT-PCR,respectively.β-actin was used as internal control.Subconfluent cells were incubated 24 and 48 hours with vari ous concentrations of Simvastatin(0.1 to 50 μmol/L),with or without 10% fetal bo vine serum.RESULTS Renal interstitial fibroblasts showe d positive-vimentin and positive-fibrobl ast surface antigen,but negative-cytoker atin.Simvastatin inhibited RFB prolifera tion in a dose-dependent manner,as compa red to the controls(P<0.05).The effect was more significant in the groups with out fetal bovine serum.The treated group s had an increased percentage of cells i n G0/G1 phase and a reduced percentage o f cells in S phase,PI values,DNA contents . But there was no significant difference between the treatment groups with resp ect to the percentage of cell in G 2/M phase.Simvastatin suppressed the express ion of c-fos mRNA in a dose-dependent ma nner as compared to the control group. CONCLUSION Simva statin suppressed RFB proliferation by i nh ibiting DNA synthesis and induceding cel l-cycle arest at G1 phase.This effect on renal interstitial fibroblasts might be exerted,at least in part,via inhibition of the c-fos and c-jun-dependent mitoge nic pathway.It implies that inhibitors o f 3-hydroxy-3-methylgutaryl coenzyme A re ductase have beneficial effect in progre ssive renal diseases.