摘要：
Objective To investigate the effects of trifluoperazine (TFP) on platelet-derived growth factor (PDGF)-induced cultured human mesangial cells (HMCs) proliferation and cell cycle.Methods (1) The measurement of cell proliferation:the HMCs of quiescent stage was divided into eight groups:control group,PDGF group,5μmol/L TFP group,10 μmol/L TFP group,15 μmol/L TFP group,20 μmol/L TFP group,25 μmol/L TFP group and 30 μmol/L TFP group before cultivated for 24 h,48 h or 72 h.MTT colorimetric method was performed to detect the levels of cell proliferation.(2) The measurement of cell cycle:the HMCs were divided into five groups:control group,PDGF group,TFP-stimulated 24 h group,TFP-stimulated 48 h group and TFP-stimulated 72 h group after quantifying,and cell cycle of HMCs was analyzed by flow cytometry.(3) The HMCs were divided into three groups:control group,PDGF group and TFP group and were cultured for 48 h.Western blot was performed to observe the instant expression of CyclinA,CDK2 and p27.Results (1) The proliferation of HMCs was increased significantly after HMCs were cultured for 24 h,48 h or 72 h with PDGF,and TFP was found to significantly inhibit the proliferation of HMCs in a dose-and time-dependent manner.(2) After using 20 μmol/L TFP,the percentage of G1 stage HMCs was significantly increased.(3) Comparing with the control group,the instant expression of CychnA and CDK2 in the PDGF group was significantly increased,while in the TFP group was decreased when compared to the PDGF group.Meanwhile,the instant expression of p27 was decreased in the PDGF group in comparison with the control group,although it was increased in the TFP group when compared with the PDGF group.Conclusions TFP can inhibit the proliferation of PDGF-induced HMCs in a dose-and timedependent manner.TFP can keep HMCs at G1 stage,which can decrease the expression of CychnA and CDK2 and increase the expression of p27.%目的 研究三氟拉嗪对血小板源生长因子诱导的人肾小球系膜细胞增殖和周期的影响.方法 (1)细胞增值测定:静止期人肾小球系膜细胞分为空白对照组、血小板源生长因子组、5μmol/L、10 μmol/L、15 μmol/L、20 μmol/L、25 μmol/L、30 μmol/L三氟拉嗪组,分别以血小板源生长因子或三氟拉嗪作用24、48、72 h,应用MTT比色法测定细胞增殖情况.(2)细胞周期测定:人肾小球系膜细胞定量后分为空白对照组、血小板源生长因子组、24h、48h、72 h三氟拉嗪组,运用流式细胞仪分析细胞周期变化.(3)人肾小球系膜细胞分为空白对照组、血小板源生长因子组、三氟拉嗪组,培养48 h后收集细胞,采用Westem blot法测定细胞周期蛋白cyclinA、CDK2、p27的表达水平.结果 (1)人肾小球系膜细胞在血小板源生长因子刺激24h、48h、72 h后增殖明显增加;三氟拉嗪可显著抑制其增殖,随着浓度和时间的增加抑制逐步明显.(2)20μmol/L三氟拉嗪作用后,人肾小球系膜细胞中G1期细胞百分数较血小板源生长因子组明显增加;随时间的延长,G1期细胞百分数逐渐增多.(3)与空白对照组比较,血小板源生长因子组周期蛋白CyclinA、CDK2表达明显增高,而三氟拉嗪组较血小板源生长因子组明显降低;周期抑制蛋白p27明显降低,而三氟拉嗪组较血小板源生长因子组明显增高.结论 三氟拉嗪能够浓度和时间依赖性地抑制人肾小球系膜细胞增殖,通过抑制CyclinA、CDK2和上调p27的表达使细胞停滞于G1期.
