缺氧诱导因子1在人肾微血管内皮细胞低温缺氧再复氧损伤中的抗凋亡作用  

Antiapoptotic Role of Hypoxia Inducible Factor 1 in Renal Micro-vascular Endothelial Cell from Cold Hypoxia/Reoxygenation Injury

作　　者：陈波[1] 赵鸿[1] 郑景存[1] 王翔[1] 瞿连喜[1] 丁强[1] 张元芳[1] 

机构地区：[1]复旦大学附属华山医院泌尿科,上海200040

出　　处：《复旦学报（医学版）》2005年第6期687-690,698,共5页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金资助项目(30200262)

摘　　要：目的探讨缺氧诱导因1(hypoxia inducible factor-1,HIF-1)在人肾微血管内皮细胞(human renal mi-crovascular endothelial cells,RMVEC)低温缺氧再复氧(hypoxia/reoxygenation,HR)损伤中的抗凋亡作用。方法建立RMVEC低温HR模型,应用3,4-DHB和NS-398预处理细胞,设对照、低温HR、3,4-DHB和NS-398共4组。采用RT-PCR和Western blot检测各组HIF-1α和血红素加氧酶1(heme oxygenases-1,HO-1)mRNA和蛋白的表达水平;通过TUNEL法及活化caspase-3蛋白表达水平检测各组细胞凋亡情况。结果各组间HIF-1αmRNA表达水平无统计学差异(P>0.05);经低温HR损伤后HIF-1α蛋白(P<0.001)、HO-1mRNA(P<0.001)和HO-1蛋白(P=0.002)表达水平与对照组比较显著增高。与低温HR组比较,3,4-DHB组HIF-1α(P<0.001)、HO-1mRNA(P=0.03)和蛋白(P=0.017)表达水平有显著增高;而NS-398组HIF-1α(P=0.036)、HO-1mRNA(P=0.007)和蛋白(P=0.005)表达水平则显著减低。低温HR损伤造成活化caspase-3表达显著增高(P=0.001)。与低温HR组比较,3,4-DHB组表达减低(P=0.005)和NS-398组表达增高(P<0.001)均有统计学意义。TUNEL结果,与对照组[(6.97±0.032)%]比较,低温HR组[(19.1±0.053)%,P=0.002]和NS-398组[(26.7±0.079)%,P<0.001]原位凋亡细胞比例显著增高;3,4-DHB组[(7.89±0.038)%]增高无统计学意义(P=0.802)。与HR组比较,3,4-DHB组减低(P=0.003)和NS-398组增高(P=0.038)均有统计学意义。结论HIF-1稳定表达在RMVEC低温HR损伤中有显著的抗凋亡作用,机制可能与其诱导内源性HO-1过表达有关。Purpose To explore the antiapoptotic role of hypoxia inducible factor-1（HIF-1） in human renal micmvascular endothelial cells （RMVEC） from hypoxia-reoxygenation （HR） injury. Methods Model of RMVEC induced by cold HR injury was established to mimic ischemia/reperfusion injury in vivo. Prolyl 4-hydroxylase inhibitor 3,4-DHB and cydooxygenase enzymes-2 blocker NS-398 were used to intervene the expression levels of HIF-la and Heme oxygenase-1（HO-1）, and the influence of them on the expression level of actived caspase-3 and the Tdt-mediated dUTP nick end-labding（TUNEL） assay. Results The expression levels of HIF-1α protein and HO-1 mRNA and protein were increased in the RMVEC pretreated with 3,4-DHB as compared with the RMVEC form cold HR injury（P〈0.01）,while the expression level of actived caspase-3 and the apopto-tic proportion were significantly decreased（P〈0.01）. The expression levels of HIF-la protein and HO-1 mRNA and protein were significantly decreased in the RMVEC pretreated with NS-398 as compared with the RMVEC from cold HR injury （P〈0.05）, the level of actived caspase-3 and the apoptotic proportion were significantly increased （P〈0.05）. Conclusions The upregulated expression of HIF-1α exerts an important antiapoptotic role in the RMVEC from cold HR injury,and this protective role may be related to it＇s ablity to induce endogenous overexpression of HO-1.

关 键 词：缺氧诱导因子1 血红素加氧酶1 低温缺氧再复氧损伤 凋亡 

分 类 号：R699.2[医药卫生—泌尿科学]