他克莫司对人肾小球系膜细胞细胞周期相关蛋白表达的影响  

The influence of tacrolimus on cyclin D1,cyclin E and p27^(kip1) in human glomereular mesangial cells

作　　者：张瑞婧[1] 李荣山[1] 白波[1] 邵珊[1] 杨鹏[1] 雒娇娇[1] 

机构地区：[1]山西医科大学第二医院肾内科 山西省肾脏病研究所,太原030013

出　　处：《中华临床医师杂志（电子版）》2012年第20期104-107,共4页Chinese Journal of Clinicians(Electronic Edition)

摘　　要：目的探讨他克莫司对体外培养的人肾小球系膜细胞细胞周期相关蛋白cyclin D1、cyclin E及p27kip1的影响及可能机制。方法设正常对照组和他克莫司处理组,他克莫司组设为三个组(浓度分别为10μmol/L、25μmol/L、50μmol/L)。四甲基偶氮唑盐法(MTT)检测他克莫司组处理12h后肾小球系膜细胞的增殖情况;流式细胞仪检测他克莫司组分别作用12、24、48h后系膜细胞的上述三种蛋白质及与其结合的CDK2和CDK4的表达情况;选择有明显抑制作用的时间点(12h),Westernblot检测上述三种蛋白质的表达情况。结果 (1)作用12h,他克莫司组均能抑制系膜细胞的增殖。(2)作用12、24、48h,与对照组相比,浓度为10μmol/L他克莫司对细胞周期蛋白cyclin D1、cyclin E及抑制因子p27kip1的表达无明显影响,而浓度为25、50μmol/L时,cyclin D1及cyclin E明显降低,p27kip1表达明显升高,呈一定的剂量效应关系;且相同浓度的他克莫司在作用三个时间点后,cyclin D1、cyclin E及p27kip1的表达无明显差异。(3)作用12h,与对照组相比,浓度为10μmol/L他克莫司对周期蛋白依赖性激酶CDK2、CDK4的表达无明显影响,而浓度为25、50μmol/L时,CDK2、CDK4的表达明显降低。结论一定浓度的他克莫司可抑制系膜细胞的增殖,其作用机制至少部分与其影响细胞周期相关蛋白的表达有关。Objective To investigate the effect of tacrulimus on the expression of cell cycle related proteins cyclin D1,cyclin E and p27kip1 in cultured human glomereular mesangial cells(HMC)and its mechanisms.Methods Cultured HMC was divided into the control group and tacrulimus treated groups of different concentrations(10 μmol/L,25 μmol/L,50 μmol/L).The proliferation of HMC was detected by MTT assay after 12 hours.The expression of cyclin D1,cyclin E and p27kip1,as well as their binding proteins CDK2 and CDK4 was measured by flow cycometry after 12,24 and 48 hours.Western blot analysis was used to assess the expression of the above proteins after 12 hours.Results Tacrolimus inhibited HMC proliferation in a concentration dependent manner.The expression of cyclin E,p27kip1,cyclin D1,CDK2 and CDK4 was inhibited by tacrolimus at the concentration of 25 and 50 μmol/L in a dose dependent manner,while tacrolimus at the concentration of 10 μmol/L had no effect.But there was no difference between different time point.Conclusions Tacrolimus inhibits the proliferation of HMC,which is achieved by,at least in part,inhibiting the expression of cell cycle related proteins.

关 键 词：他罗利姆 细胞周期蛋白D1 细胞周期蛋白E 细胞周期 肾小球系膜细胞 p27~kip1 

分 类 号：R692[医药卫生—泌尿科学]