Akt3基因在低氧致大鼠肺动脉平滑肌细胞体外增殖中的作用  

The role of Akt3 gene in pulmonary arterial smooth muscle cell in vitro amplication induced by hypoxia in rats

作　　者：易斌[1] 钱桂生[1] 白莉[1] 王关嵩[1] 

机构地区：[1]第三军医大学新桥医院全军呼吸研究所,重庆400037

出　　处：《重庆医学》2008年第22期2553-2555,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(30400200;30700347);重庆市自然科学基金资助项目(2007BB5045)

摘　　要：目的通过观察缺氧致大鼠肺动脉平滑肌细胞(PASMC)丝氨酸/苏氨酸蛋白激酶3(Akt3)mRNA及其蛋白表达水平变化与低氧PASMC增殖的关系,探讨Akt3在低氧肺血管重建中的调控作用。方法组织块法培养PASMC。采用半定量RT-PCR技术检测常氧组(N组)和低氧2、8、12、24h组Akt3基因mRNA表达水平;采用Westernblot技术检测相应蛋白表达水平。采用MTT法、3H-TdR掺入法检测PASMCs增殖改变。结果低氧诱导PASMCs不断增殖。各组均检测出Akt3基因mRNA以及蛋白表达变化,图像定量分析光密度值显示各组表达mRNA以及蛋白表达与细胞增殖改变密切相关。结论Akt3基因可能在低氧肺动脉高压中调节PASMC增殖。Objective To observe the changes of Akt3 mRNA and protein expressive level in pulmonary arterial smooth muscle cells （PASMC） induced by hypoxia in rats,and to investigate the regulatory role of Akt3 signaling pathway in hypoxia-induced pulmonary vascular remodeling. Methods To establish the pure PASMCs culture system by tissue-sticking method. Semi-quantitative reverse transcription and polymerase chain reaction （RT-PCR） were used to examine the Akt3 mRNA expression after PASMCs were exposed to hypoxia for 2h,8h, 12h and 24h respectively. And PTEN protein expressions were determined by Western blotting. The changes of PASMCs proliferation were determined by MTT and 3 H-TdR incorporated way. Results Hypoxia could stimulate PASMCs to proliferate uninterruptedly. The mRNA and protein of Akt3 could be detected in all groups,and the expression level of mRNA and protein were closely associated with PASMCs proliferation. Conclusion Akt3 signaling pathway may play an important role in regulating PASMC proliferation in hypoxia-induced pulmonary hypertension.

关 键 词：平滑肌细胞 丝氨酸/苏氨酸蛋白激酶 缺氧肺动脉高压 

分 类 号：R363[医药卫生—病理学]