缺氧-复氧肾小管上皮细胞损伤的信号传导机制

Progression on signal transduction pathway mechanism of HK-2 cells injury induced by hypoxia-reoxygenation

目的探讨肾小管上皮细胞在缺氧-复氧(H-R)损伤后的信号传导机制。方法将不同方法处理的人近曲肾小管上皮细胞(HK-2)分为对照组、模型组和PDTC组,模型组和PDTC组再分别分成缺氧6、12、24h组、缺氧24h后分别复氧6、12、24h组;检测细胞成活率、NF-κB p65蛋白表达、细胞间黏附分子-1(ICAM-1)mRNA及蛋白表达情况。结果与对照组比较,模型组随缺氧时间延长细胞数量逐渐减少,24h达高峰(P<0.05),复氧后细胞数量略有增加;随缺氧时间延长NF-κB p65阳性蛋白表达增多,复氧6h组达高峰(P<0.05);ICAM-1mRNA及蛋白表达随H-R时间延长渐增高(P<0.05)。PDTC组中NF-κB p65蛋白和ICAM-1mRNA及蛋白表达较模型组有明显下降(P<0.05)。结论 H-R诱导HK-2细胞中NF-κB p65的活化,从而上调ICAM-1mRNA及蛋白表达。

Objective To investigate the signal transduction pathway mechanism of human renal tubular epithelial cell damage induced by hypoxia-reoxygenation. Methods Human renal tubular epithelial cell line HK-2 cell was used as target cell. The experiment are divided into the control group, the model group and PDTC group. Each group was further divided into 6 subgroups according to the times of hypoxia or/and reperfusion（H6, H12, H24,R6 ,R12 and R24）. Detected the cell survival, the protein expres- sion of nuclear factor-kB（NF-kB）protein65, the mRNA and protein expression of ICAM-1 respectively. Results Compare with control group, cell survial were significantly decreased and droped the lowest in group H24（P〈0.05）which induced by Hypoxia. H/R up-regulates NF-kB protein65,and the mRNA and protein expression of ICAM-1 of the cell increased significantly（P〈0.05）. In addition,expression of NF-kB protein65 and ICAM-1 which induced by H-R were remarkably reduced in the PDTC group（P〈0. 05）. Conclusion H/R induced NF-kB activation and increased ICAM-I mRNA and protein expression,this effect can be inhibit by PDTC. It suggested that H/R induces ICAM-1 mRNA and protein expression in HK-2 cells through activation of NFkB.