中文摘要：

目的研究抑制整合素连接激酶（ILK）表达对大鼠肾小球系膜细胞（RMC）细胞间隙连接蛋白43（Cx43）表达的影响。方法将大鼠肾小球系膜细胞分为RMC组（n-6）、ILK-consiRNA转染组（n=6）和ILK-siRNA转染组（n=6）。合成抑制ILK基因的siRNA，待细胞长至60％融合后，ILK-siRNA转染组和ILK-con siRNA转染组分别用脂质体转染ILK-siRNA或ILK-con siRNA，RMC组仅加入脂质体，24h后收集细胞，提取总蛋白和总RNA，用RT-PCR和Western blotting观察ILK和Cx43 mRNA和蛋白的表达。转染后继续培养24、48、72h，MTT法检测细胞活力。结果与RMC组和ILK-con siRNA转染组比较，ILK-siRNA组ILK mRNA和蛋白的表达均下降30％-50％（P〈0.05，P〈0.01），Cx43 mRNA水平增加30％-40％（P〈0.05），Cx43蛋白水平增加60％-70％（P〈0.01）。应用ILK-siRNA转染系膜细胞后24、48、72h，细胞活力均显著高于RMC组和ILK-con siRNA转染组（P〈0.05，P〈0.01）。结论抑制ILK途径可上调肾小球系膜细胞Cx43的表达，增强细胞活力；Cx43的调节可能是部分通过ILK途径实现的。

英文摘要：

Objective To investigate the effects of the expression of integerin-linked kinase（ILK） on connexin 43（Cx 43） in rat mesangial cells （RMCs）. Methods RMCs were divided into three different groups （6 for each group）：RMC group, ILK-con siRNA group and ILK-siRNA group. ILK siRNA was synthesized, and then transfected into RMCs by LipofectAMIN 2000. RMCs were transi- ently tranefected with ILK-con siRNA, ILK-siRNA and lysed 24h later, and mRNA was then extracted and detected by reverse transcription-polymerase chain reaction（RT-PCR）analysis using ILK and Cx43 specific primers, and an aliquot of protein from each sample was subjected to western blot analysis using ILK and Cx43 antibodies. Cells were seeded into 96-well plates （2×10^3 cells/well） and then trans-fected with ILK con siRNA and ILK siRNA. After incubation for 24, 48 and 72 hours, respectively, 20μl of 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT）（5mg/ml） was added into each well and incubated for 4 hours. Subsequently, 150μl of dimethyl sulfoxide （DMSO） was added to each well to dissolve the forrnazan crystals, and the absorption at 492nm was measured. Results ILK mRNA and protein levels decreased by 30%-50% after being transiently transfeeted with ILK-siRNA, while Cx43 mRNA and protein levels increased by 30%-40% and 60%-70%, respectively. The viabilities in ILK-siRNA infected RMCs at 24, 48 and 72 hours were significantly higher than that in ILK con siRNA infected RMCs. Condusion Inhibition of ILK pathway will up-regulate the expression of Cx 43 and the viability of RMCs, implying that the regulation of connexin 43 might possibly be achieved via ILK pathway.