舒拉明对血清刺激的大鼠肾间质成纤维细胞增殖的研究

Study of Suramin on Rats Interstitial Fibroblasts Proliferation

目的:肾间质成纤维细胞增殖是肾间质纤维化的重要启动机制之一,本研究将探讨舒拉明对血清刺激的大鼠肾间质成纤维细胞增殖的影响及其调控机制。方法:体外培养大鼠肾间质成纤维细胞(NRK-49F),舒拉明以不同浓度(0,10,50,100μmol/L)处理细胞,观察细胞数目变化,显微镜拍照,MTT方法检测细胞活性变化,免疫印迹技术检测细胞周期蛋白Cyclin D1和P27的变化。结果:研究发现,舒拉明以剂量依赖性效应抑制细胞生长;MTT方法检测发现随着舒拉明剂量的加大,显著下调细胞增殖活性;通过检测细胞周期正向调控蛋白(Cyclin D1)和细胞周期负向调控蛋白(P27),证实舒拉明可剂量依赖性效应抑制Cyclin D1表达,增强细胞周期负向调控蛋白P27表达。结论:舒拉明可在体外以剂量依赖性效应抑制肾间质成纤维细胞增殖,可能与其调控细胞周期蛋白表达有关。

Objective： Massive proliferation of renal interstitial fibroblasts has been considered as one of the most pivotal and fundamental mechanisms in renal interstitial fibrosis. Our study aims to elucidate the effects of Suramin on serum- stimulated rats renal interstitial fibroblast proliferation and mechanisms involved. Methods： Cultured rat renal interstitial fibroblasts（ NRK- 49F） were treated with Suramin at different doses（ 0,10,50,100 μmol / L）. Cell proliferation was assessed by counting cells,the MTT（ 3-（ 4,5- dimethylthiazol- 2- yl）- 2,5- diphenyltetrazolium bromide） assay. Expression of Cyclin D1,a positive cell cycle regulator,and expression of p27,a negative cell cycle regulator were also detected by western blot. Results： Our study showed that Suramin blocked cell growth in dose- dependent effect. Suramin inhibits cell proliferation activity significantly by MTT assay. Furthermore,suramin decreased expression of Cyclin D1,and increased expression of p27 in dose- dependant effection. Conclusion： Suramin can block proliferation of cultured renal interstitial fibroblasts in dose- dependent manner,which is possibly connected with regulation of cell cycle protein expression by suramin.