缺氧诱导因子-1对肾微血管内皮细胞缺氧再复氧损伤免疫调节作用  

Protective immuofunction of hypoxia inducible factor-1 on microvascular endothelial cell with hypoxia-reoxygenation

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外科,上海200040

出　　处：《中华实验外科杂志》2005年第9期1043-1045,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30200262)

摘　　要：目的探讨低氧诱导因子1(HIF1)在肾微血管内皮细胞(HGMECs)缺氧再复氧损伤中的免疫调节作用。方法建立HGMECs缺氧再复氧模型,应用特异性脯氨酰羟化酶抑制剂3,4DHB和环氧酶2阻断药物NS398预处理HGMECs,设对照、缺氧复氧、3,4DHB和NS398四组。采用逆转录聚合酶链反应、免疫印记和酶联免疫吸附试验等方法检测各组细胞HIF1α、血红素氧合酶1(HO1)的mRNA和蛋白的表达及上清中可溶性细胞间黏附因子1(sICAM1)的水平;将各组HGMECs与异体淋巴细胞混合培养(MELR),检测淋巴细胞增殖程度和MELR上清中白细胞介素2(IL2)的表达水平。结果与对照组相比,HGMECs经缺氧复氧损伤后HIF1α蛋白和HO1mRNA表达水平显著增高(P<0.01),上清中sICAM1(14.55±1.13vs7.90±1.61)ng、异体淋巴细胞增殖率(0.191±0.053vs0.069±0.032)和MELR上清中IL2(40.0±5.0vs18.0±3.0)pg水平均有显著增高(P<0.01)。与缺氧复氧组比较,3,4DHB干预组HIF1α蛋白和HO1mRNA表达水平显著增高(P<0.05),而淋巴细胞增殖率(0.079±0.038,P<0.01)及IL2(25.7±6.1pg,P<0.01)和sICAM1(8.71±1.32ng,P<0.01)水平则显著下调。与缺氧复氧组比较,NS398干预组HIF1α蛋白和HO1mRNA表达水平显著减低(P<0.01),而淋巴细胞增殖率(0.268±0.079)及IL2(51.0±11.0)pg和sICAM1(20.33±3.05)ng表达水平则显著增高(P<0.05)。结论缺氧复氧会对HGMECs造成免疫学损伤,HIF1α稳定表达可起到保护性作用。Objective To explore the influence of hypoxia inducible factor-1 （HIF-1） on the immunofunction of human glomerular microvascular endothelial cells （HGMECs） with hypoxia-reoxygenation （HR）. Methods Model of HGMECs induced by HR was established to mimic ischemia/reperfusion injury. Prolyl 4-hydroxylase inhibitor 3, 4-DHB and cyclooxygenase enzymes-2 （COX-2） blocker NS-398 were used to intervene the expression of HIF-1 and heme oxygenase-1 （HO-1）. Proliferative rate of lym- phocytes stimulated by the HGMECs was measured by MTT test, while the supematant levels of IL-2 and ICAM-1 were detected by ELISA. Results As compared with HGMECSs with FIR, the expression of HIF-la protein and HO-1 mRNA was significantly increased in HGMECs pretreated with 3, 4-DHB （ P〈0.05 ）. While the supematant levels of IL-2 [ （25.7±6.1 ） vs （40.0±5.0） pg, P〈0.01 ], ICAM-1 [ （8.71±1.32） vs （14.55±1.13） ng, P〈0.01] and the lymphocyte proliferative rate （0. 079±0. 038 vs 0. 191±0. 053, P=0. 003） were significantly decreased. On the contrary, the expression of HIF-la protein and HO-1 mRNA was markedly decreased in the HGMECs pretreated with NS-398 as compared with the HGMECs with HR （P〈0.05）. Conclusion The overexpression of HIF-la may have protective immunofunction on the HGMECs with HR.

关 键 词：血管内皮细胞 缺氧诱导因子1 血红素加氧酶1 缺氧 缺氧再复氧损伤 微血管内皮细胞 免疫调节作用 缺氧诱导因子-1 逆转录-聚合酶链反应 淋巴细胞增殖 

分 类 号：R692[医药卫生—泌尿科学]