不同程度低氧对大鼠肺动脉平滑肌细胞增殖和HIF-1αmRNA表达的影响  

EFFECTS ON HIF-1 ACTIVITY AND PROLIFERATION OF RAT PULMONARY ARTERIAL SMOOTH MUSCLE CELLS IN DIFFERENT HYPOXIA

作　　者：张昱[1] 黄以哲 刘瑞欣[1] 张伟[1] 

机构地区：[1]青海大学医学院 [2]青海省武警总队医院

出　　处：《青海医学院学报》2013年第2期73-78,共6页Journal of Qinghai Medical College

基　　金：国家自然基金项目(81160012);教育部"春晖计划"科技项目(Z2005-2-81003)

摘　　要：目的探讨不同低氧环境对大鼠肺动脉平滑肌细胞(PASMCs)增殖和HIF-1αmRNA表达的影响。方法采用原代培养大鼠PASMCs,分别在常氧、低氧(10%O2,85%N2,5%CO2)和缺氧(1%O2,94%N2,5%CO2)条件下进行培养,应用四甲基偶氮唑蓝(MTT)比色法检测细胞的增殖水平,通过不同低氧环境培养48 h后,用RT-PCR检测PASMCs HIF-1αmRNA表达水平。结果连续7 d MTT监测结果表明,低氧组细胞的生长和常氧组相比无显著性差异。缺氧组的PASMCs 2 d后明显增殖,与常氧组相比有显著性差异(P<0.01),5 d后细胞数达到顶峰,6 d后缺氧组细胞开始出现空泡化和死亡,细胞数开始减少。不同低氧条件下PASMCs培养48 h后,低氧组PASMCs的HIF-1αmRNA表达水平和常氧组相比没有显著性差异,缺氧组PASMCsHIF-1αmRNA表达水平显著升高(P<0.01)。结论 HIF-1α是PASMCs低氧增殖的关键因子,在低氧环境下且造成PASMCs的"缺氧"后才能使细胞HIF-1αmRNA表达升高,促进细胞增殖。Objective To explore the effects on HIF - 1α mRNA expressions and proliferation of rat pulmonary arterial smooth muscle ceils （PASMCs）in different hypoxia. Methods The PASMCs of rat had been isolated and cul- tured. Then the PASMCs were cultured in normoxia（21% 02 and 5% CO2） ,hypoxia（ 10% 02,5% CO2 and 85 % N2 ） and anoxia（ 1% 02 ,5% CO2 and 94 % N2 ） condition respectively. The cell proliferation was tested by methyl thiazolyl tetrazolium （MTY） method. After 48 hrs in different hypoxia, the mRNA expression of HIF - 1α was detec- ted by RT - PCR. Results After 7 days of detection, the proliferations of PASMCs in Normoxia group and Hypoxia group were similar. The proliferation of PASMCs in Anoxia group was stronger than Normoxia group and Hypoxia group after 2 days ,and the cell number reached the peak on 5th day. The PASMCs of Anoxia group occurred vacuol-ization and death from 6 days. After 48 hrs in different hypoxia,the expression of PASMCs＇ HIF - 1α mRNA of the Normoxia group and Hypoxia group are not increased. In Anoxia group,the expression of PASMCs＇ HIF- 1α mRNA was increased remarkably. Conclusion These results indicate that HIF - 1α plays a pivotal role in PASMCs prolifer- ation. It may stimulate HIF - 1 mRNA express of PASMCs in hypoxia, and promote proliferation.

关 键 词：缺氧 肺动脉平滑肌细胞 缺氧诱导因子-1Α 细胞增殖 

分 类 号：R322.121[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]