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目的:研究重组腺病毒介导的人胞质型谷胱甘肽过氧化物酶(hCGPx)转染对人肾小管上皮细胞缺氧再复氧损伤的保护作用。方法:将含hCGPXcDNA的质粒pGEM-T-hCG-Px和重组腺病毒穿梭质粒pACCMV-pLpA重组,构建pACC-MV-hCGPx穿梭质粒后,与包装质粒pJM17共转染293细胞,构建重组腺病毒AdCMV-hCGPx。以重组腺病毒载体AdCMV-hCGPx转染体外培养的人肾小管上皮细胞株HK-2,以转染空载体的HK-2细胞为对照组,检测转染细胞中CGPx的表达水平。将HK-2细胞经缺氧再复氧损伤处理后,分别检测细胞的存活率、凋亡率及死亡率。结果:各转染组细胞中CGPx的表达率显著高于对照组(P<0.01)。经缺氧再复氧损伤处理后,AdCMV-hCGPx转染组细胞的存活率较对照组明显增强,死亡细胞明显减少,细胞凋亡明显受到抑制。结论:重组腺病毒介导的hCGPx转染人肾小管上皮细胞可保护缺氧再复氧引起的损伤。
AIM: To investigate the protective effect of recombinant adenovirus mediated transfection of hCGPx on hypoxia and reoxygenation injury of human renal tubular epithelial cells. Methods: The recombinant plasmid pGEM-T-hCG-Px containing hCGPXcDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were recombined to construct the shuttle plasmid pACC-MV-hCGPx. The recombinant plasmid pCMV-hCGPx was co-transfected with the plasmid pJM17 into 293 cells to construct the recombinant adenovirus AdCMV -hCGPx. The human renal tubular epithelial cell line HK-2 cultured in vitro was transfected with the recombinant adenovirus vector AdCMV-hCGPx. The transfected cells were transfected with empty vector of HK-2 cells as the control group to detect the expression level of CGPx. HK-2 cells were treated with hypoxia and reoxygenation, and the cell viability, apoptosis rate and mortality were detected. Results: The expression of CGPx in each transfected group was significantly higher than that in control group (P <0.01). After hypoxia and reoxygenation treatment, the survival rate of AdCMV-hCGPx transfection group was significantly higher than that of control group, the number of dead cells was significantly decreased, and the apoptosis was obviously inhibited. CONCLUSION: Recombinant adenovirus-mediated hCGPx transfected human renal tubular epithelial cells protects against hypoxia-reoxygenation-induced injury.