丹参酮ⅡA磺酸钠对肾间质成纤维细胞P16蛋白表达的影响

Effect of DS- 201 on P16 expression in human renal interstitial fibroblasts

目的:观察丹参酮ⅡA磺酸钠(sodium tanshinoneⅡA sulfonate,DS-201)对人肾间质成纤维细胞(human renal interstitial fibroblasts,hRIFs)体外增殖以及P16表达的影响,试阐明其在延缓肾间质纤维化过程中作用的具体环节,为中药丹参治疗肾间质纤维化(renal interstitial fibrosis,RIF)提供理论和实验依据。方法:用四甲基偶氮唑(MTT)法检测对照组与不同DS-201浓度组hRIFs的增殖活性。免疫细胞化学技术检测0 mg/ml、0.5 mg/ml、1.0 mg/ml、2.0 mg/ml DS-201孵育hRIFs 72 h后P16的表达。结果:与对照组相比,各DS-201组hRIFs细胞由均一的梭形变成椭圆形甚至圆形;随着用药浓度增高,hRIFs抑制率明显增高;P16表达增强,平均光密度值显著增加,组间差异有统计学意义(P<0.05)。结论:DS-201可能通过促进P16的表达从而抑制hRIFs通过G1/S关卡,使细胞阻滞于G0/G1期,进而抑制细胞增殖。

Objective： To observe the effect of sodium tanshinone ⅡA sulfonate（DS - 201） on proliferation activity of human renal interstitial fibroblasts and P16 expression in vitro. In order to approach its molecular mechanism delaying the renal interstitial fibrosis. Methods： The proliferation of hRIFs cultured in 3 different concentrations of DS-201 was tested by MTT analysis and the expression of P16 was tested with immunocytochemical method. Results： In DS - 201 groups, the proliferation activity of hRIFs was inhibited（P﹤0.05）and the expression of P16 was up-regulated（P﹤0.05）. Conclusion： The inhibitory effects of DS - 201 on hRIFs proliferation might be due to the stimulation of P16 expression, by which cells were arrested at G0 / Gl phase.