盐酸吡格列酮对肾小球系膜细胞megsin基因表达和α-SMA、TGF-β_1合成的影响  

Effects of pioglitazone hydrochloride on expression of megsin gene and syntheses of TGF-β_1 and α-SMA in human mesangial cells

作　　者：李霞[1] 徐佳岱[2] 郭德森[3] 戴春[1] 

机构地区：[1]徐州医学院附属医院肾脏内科,221000 [2]南京医科大学基础医学院 [3]内蒙古民族大学附属医院

出　　处：《江苏医药》2009年第4期452-454,I0002,共4页Jiangsu Medical Journal

摘　　要：目的研究盐酸吡格列酮对培养的人肾小球系膜细胞(hMC)megsin基因表达及α平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)合成的影响。方法培养hMC,设计并合成针对megsin基因编码区的siRNA双链,经原位杂交方法筛选出有效的序列,采用siRNA专用脂质体RNA-mate将siRNA转染至hMC,同时设盐酸吡格列酮组(1、5、10μmol/L三种浓度)和正常对照组。MTT方法检测细胞增殖,免疫组化方法检测α-SMA、TGF-β1的合成,原位杂交法检测细胞megsin mRNA表达。结果不同浓度盐酸吡格列酮可以抑制hMC megsin基因mRNA的表达,48 h抑制效果最强。megsin siRNA能够阻抑hMC细胞megsin基因表达。结论盐酸吡格列酮可以抑制hMC megsin基因mRNA的表达,但对体外培养的hMC增值无明显影响,对α-SMA、TGF-β1合成无明显影响。Objective To investigate the effects of pioglitazone hydrochloride on the expression of megsin gene and syntheses of transforming growth factor-β1 （TGF-β1） and a-smooth muscle acting protein （α-SMA） in human mesangial cell（hMC）. Methods hMC was cultured in vitro, siRNA double strands targeting megsin coding region was designed and synthesized. An effective sequence was selected by in-situ hybridization, and transfected into human mesangial cells by RNA-mate. Different concentrations（1, 5 and 5 mol/L） of pioglitazone hydrochlorid were used in the experiment groups and control group. MTT assay was used for detecting the cell proliferation. Immunohistochemical staining was conducted to investigate the effects of siRNA and pioglitazone hydrochlorid on synthesis of TGF-β1 and α-SMA. In-situ hybridization was used to detect the expression of megsin m-RNA. Results Three concentrations of pioglitazone hydrochlorid inhibited the expression of hMC megsin mRNA with the peak effect at 48 h. Megsin siRNA inhibited the expression of megsin mRNA. Conclusion Pioglitazone hydrochlorid can inhibit the expression of megsin mRNA without effect on the hMC proliferation in vitro and on the synthesis of α-SMA and TGF-β1.

关 键 词：盐酸吡格列酮 人肾小球系膜细胞 Megsin基因 Α平滑肌肌动蛋白 转化生长因子岛 

分 类 号：R575[医药卫生—消化系统]