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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 the synthesis of α-SMA and TGF-β1 in cultured human glomerular mesangial cells (hMCs). Methods hMCs were designed and synthesized. The siRNA double strands against the coding region of megsin gene were designed and synthesized. The effective sequences were screened by in situ hybridization. The siRNA was transfected into hMC by liposome RNA-mate. Meanwhile, pioglitazone hydrochloride group 1,5,10μmol / L three concentrations) and normal control group. The cell proliferation was detected by MTT assay. The expression of α-SMA and TGF-β1 was detected by immunohistochemistry. The expression of megsin mRNA was detected by in situ hybridization. Results Pioglitazone hydrochloride could inhibit the mRNA expression of hMC megsin gene at 48 h. Megsin siRNA represses megsin gene expression in hMC cells. Conclusion Pioglitazone hydrochloride can inhibit the mRNA expression of hMC megsin gene, but has no significant effect on the proliferation of hMC cultured in vitro, but has no significant effect on the synthesis of α-SMA and TGF-β1.