缺氧诱导因子-1α基因转染对缺氧损伤HepG2细胞的保护作用  

Protective effects of HIF-1α gene transfection against hypoxic injury in HepG2 cells

作　　者：蒋春华[1,2,3] 罗勇军[2,3,4] 黄庆愿[1,2,3] 高钰琪[1,2,3] 

机构地区：[1]第三军医大学高原军事医学系病理生理学与高原生理学教研室,重庆400038 [2]高原医学教育部重点实验室,重庆400038 [3]全军高原生理与高原病研究重点实验室,重庆400038 [4]第三军医大学高原军事医学系高原疾病学教研室,重庆400038

出　　处：《中国病理生理杂志》2010年第1期1-6,共6页Chinese Journal of Pathophysiology

基　　金：国家重点基础研究发展计划(973)资助项目(No.2006CB504101);国家自然科学基金面上资助项目(No.30771043)

摘　　要：目的:观察缺氧诱导因子-1α(HIF-1α)基因转染对HepG2细胞缺氧损伤的影响并初步探讨其作用机制。方法:腺病毒转染的人HepG2细胞常氧培养24h后分为4组:Ad-GFP转染常氧培养组(Ad-GFP transfected-normoxia组)、Ad-HIF转染常氧培养组(Ad-HIF transfected-normoxia组)、Ad-GFP转染低氧培养组(Ad-GFP transfected-hypoxia组)和Ad-HIF转染低氧培养组(Ad-HIF transfected-hypoxia组)。观察各组细胞的细胞活力、乳酸脱氢酶(LDH)释放率、细胞内活性氧(ROS)、一氧化氮(NO)含量和诱导型一氧化氮合酶(iN-OS)活力。结果:Ad-HIF转染的HepG2细胞可高效表达HIF-1α。Ad-GFP transfected-hypoxia组的细胞活力显著低于Ad-GFP transfected-normoxia组(P<0.05);Ad-HIF transfected-hypoxia组的细胞活力与Ad-HIF transfected-normoxia组比较无显著差异。Ad-GFP transfected-hypoxia组细胞内ROS含量显著高于Ad-GFP transfected-normoxia组(P<0.05);Ad-HIF transfected-hypoxia细胞内ROS含量与Ad-HIF transfected-normox-ia组或Ad-GFP transfected-hypoxia组比较均无显著差异。Ad-HIF transfected-normoxia组和Ad-GFP trans-fected-hypoxia组细胞内NO含量和iNOS活力均显著高于Ad-GFP transfected-normoxia组(P<0.05);Ad-HIFtransfected-hypoxia组细胞内NO含量和iNOS活力与Ad-GFP transfected-hypoxia组和Ad-HIF transfected-nor-moxia组比较无显著差异。结论:基础性HIF-1高表达可显著减轻缺氧时HepG2细胞的损伤程度,其机制可能与HIF-1高表达提高HIF-1调控的缺氧相关基因的基础表达水平和其产物含量有关;也可能与HIF-1高表达防止缺氧时ROS的过度增加有关。AIM： To study the protective effects of HIF -1α gene transfeetion on hypoxic injury in human HepG2 cells. METHODS： After gene transfection, HepG2 cells were randomly divided into 4 groups ： normoxia with Ad - GFP transfected group, normoxia with Ad - HIF - 1 transfected group, hypoxia with Ad - GFP transfected group and hypoxia with Ad - HIF - 1 transfeeted group. LDH leaking rate, cell viability, contents of NO and ROS, the iNOS activity were measured. RESULTS： High levels of HIF -1α mRNA and protein were detected in Ad - HIF transfected HepG2 cells. Cell viability was significantly lower in Ad - GFP transfected - hypoxia group than that in Ad - GFP transfeeted - normoxia group （ P 〈 0. 05 ）. No marked difference of cell viability was found between Ad - HIF transfected - hypoxia group and Ad - HIF transfected - normoxia group. ROS was significantly higher in Ad - GFP transfected - hypoxia group than that in Ad - GFP transfected - normoxia group （ P 〈 0. 05 ）, while no marked difference was found either between Ad - HIF transfected - hypoxia group and Ad - HIF transfected - normoxia group or between Ad - HIF transfected - hypoxia group and Ad - GFP transfected - hypoxia group. The content of NO and iNOS activity were significantly higher in Ad - HIF transfected - normoxia group and Ad - GFP transfected - hypoxia group than those in Ad - GFP transfected - normoxia group （ P 〈0. 05 ）, no marked difference was found either between Ad - HIF transfected - hypoxia group and Ad - GFP transfected - hypoxia group or between Ad - HIF transfected - hypoxia group and Ad - HIF transfected - normoxia group. CONCLUSION ： Higher HIF -1α expression is contributed to protective effects against hypoxic injury in HepG2 cells, the mechanisms of which may be correlated with promoting expression of gene regulated by HIF - 1 and restraining over - expression of injure factors.

关 键 词：缺氧 缺氧诱导因子-1 基因转染 腺病毒载体 

分 类 号：R363[医药卫生—病理学]