低氧诱导因子1在低氧致肺动脉平滑肌细胞增殖中的作用  

Effects of hypoxia-inducible factor-1 signal pathway on proliferation in hypoxic pulmonary artery smooth muscle cells

作　　者：李炽观[1] 戴爱国 严鹏科[3] 

机构地区：[1]郴州市第一人民医院呼吸内科,湖南郴州423000 [2]湖南省老年医院湖南省老年医学研究所呼吸病研究室,湖南长沙410001 [3]南华大学药理研究所,湖南衡阳421001

出　　处：《中国病理生理杂志》2007年第7期1301-1305,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No30270581);湖南省教育厅重点科研基金资助项目(No02A047);湖南省自然科学基金资助项目(No99JJY20033)

摘　　要：目的:探讨低氧对肺动脉平滑肌细胞(PASMC)增殖和凋亡的影响,以及HIF-1α、P-ERK1/2、iN-OS蛋白表达变化在其中的作用与意义。方法:体外培养大鼠PASMC,设计常氧组、低氧组及ADM、L-NAME、PD98059干预组,用MTT比色法和PCNA的免疫组化法测定细胞增殖反应,用流式细胞仪检测细胞凋亡,用West-ern blotting法检测HIF-1α、P-ERK1/2、iNOS的蛋白表达。结果:(1)低氧24h组的A值明显高于常氧组(P<0.01),而PD98059及ADM干预组明显低于低氧组(P<0.01),L-NAME干预组明显高于低氧组和常氧组(P<0.01)。(2)免疫组化表明,低氧24h组呈阳性表达(P<0.01)。PD98059、ADM抑制了PCNA的表达(P<0.01),L-NAME促进了PCNA的表达(P<0.01)。(3)各组在低氧培养24h后,凋亡指数差异无显著(均P>0.05)。(4)Western blotting表明常氧组少量HIF-1α、iNOS、P-ERK1/2表达,低氧4h后均表达增高(P<0.01),8h仍维持在高峰(P<0.01),而HIF-1α、P-ERK1/2在低氧24h后表达下调。L-NAME促进了HIF-1α表达(P<0.01),PD98059部分抑制了HIF-1α、iNOS及P-ERK1/2表达(P<0.01);ADM部分抑制了HIF-1α表达,促进iNOS表达(P<0.01)。结论:低氧能促进肺动脉平滑肌细胞增殖,对细胞的凋亡无影响;HIF-1在低氧诱导肺动脉平滑肌细胞增殖中起重要作用。AIM： To investigate the effect of hypoxia on the proliferation and apoptosis of pulmonary artery smooth muscle cells （PASMC） ; and to evaluate the role of hypoxia - inducible factor - 1α（ HIF - 1α） , iNOS, P - ERK1/2 protein expression in hypoxie pulmonary hypertension （HPH） pathogenesis. METHODS ： Cultured rat PASMC were divided into normoxie group; hypoxie group; hypoxia ＋ ADM （adrenomedulin） group, hypoxia ＋ L - NAME （iNOS inhibitor） group ; hypoxia ＋ PD98059 group. Proliferation was investigated by MTT and PCNA. Apoptosis was examined by flow - cytometry. Westen blotting was used to measure protein expression of HIF -1α, P - ERK1/2 and iNOS. RESULTS： （ 1 ） A value of 24 h - hypoxia was significantly higher than that in the normoxie group （ P 〈 0. 01 ）. In the hypoxia ＋ PD98059 group, ADM was significantly lower than that in hypoxia group, whereas A value of the hypoxia ＋ L - NAME was significantly higher than that in hypoxie group and normoxie group （P 〈0. 01 ）. （2） PCNA was positive in PASMC after 24 h hypoxia （P 〈0. 01 ）. PD98059, ADM inhibited the expression of PCNA significantly （P 〈0. 01 ） , whereas L -NAME increased the expression of PCNA significantly （P 〈 0.01 ）. （3） Apoptosis index was not significantly difference among the different groups （P 〉0. 05）. （4） HIF -1α, iNOS and P - ERK1L2 expression was poorly positive in normoxie group, positive after hypoxia for4h （P〈0. 01 ）, reaching its peak at 8 h hypoxia （P〈0. 01 ）, HIF -1α, P - ERK1/2 expression dedined after 24 h hypoxia. L - NAME promoted the expression of HIF -1α, PD98059 inhibited the expression of HIF -1α, iNOS and P - ERK1/2 partly. ADM inhibited the expression of HIF -1α partly, promoted the expression of iNOS. CON- CLUSION ： （ 1 ） Hypoxia stimulates the proliferation of PASMC, and has no obvious effects on the apoptosis of PASMC. （ 2 ） HIF - 1 plays an importent role in the proliferation of hvpoxie PASMC.

关 键 词：缺氧诱导因子1 平滑肌细胞 肺动脉 一氧化氮合酶 缺氧 

分 类 号：R363[医药卫生—病理学]