缺氧致大鼠肺动脉平滑肌细胞Akt mRNA表达的变化

Changes of Akt mRNA expression in rat pulmonary arterial smooth muscle cells induced by hypoxia

目的通过观察缺氧致大鼠肺动脉平滑肌细胞(pu lmonary arterial smooth musc le cell,PASMC)3种丝氨酸/苏氨酸蛋白激酶(Akt1、Akt2、Akt3)mRNA的表达水平变化,初步探讨Akt信号途径在缺氧肺动脉高压(hypoxia pu lmonaryhypertension,HPH)血管重建中的作用。方法组织贴片法建立纯PASMC细胞系。采用半定量RT-PCR技术检测常氧组(N组)、低氧2、8、12、24组的Akt1、Akt2和Akt3基因mRNA的表达水平。结果建立了纯PASMC细胞系。各组均检测出Akt1、Akt2和Akt3基因的mRNA,图像定量分析光值显示各组表达的mRNA含量与缺氧时限存在一定的关系。结论Akt信号通路可能是大鼠缺氧肺动脉高压中PASMC增殖和分化的重要传导途径,其中Akt1可能起主要作用。

Objective To observe the changes of Aktl, Akt2,Akt3 mRNA expression level in rat pulmonary arterial smooth muscle cells （PASMC） induced by hypoxia and to investigate the value of Akt signaling pathway in vascular reconstruction during phyoxia pulmonary hypertension （HPH）. Methods The pure PASMCs were established by tissue-sticking methods. Semi-quantitative reverse transcription polymerase chain reaction （RT-PCR） were used to examine the Aktl ,Akt2 and Akt3 mRNA expression after PASMCs were exposed to normoxia （21% O2） or hypoxia （3%O2 ） for 2, 8, 12 and 24 h respectively. Results The pure PASMCs were established. The mRNA of Aktl ,Akt2 ,Akt3 could be detected in all groups. Exposed to hypoxia, the mRNA expression levels gradually increased, reached the peak at 8 h, then decreased, and at 24 h down to the level under normoxia. As compared with that exposed to normoxia, the mRNA expression levels of Aktl at 2, 8, 12, 24 h, Akt2 at 8 h, Akt3 at 8 h in PASMCs exposed to hypoxia showed significant difference （P 〈 0. 05）. There was a significant difference in the mRNA expression of Aktl, Akt2, Akt3 between at 2 h and 8 h, as well as Aktl at 12, 24 h, Akt2 at 12 h, Akt3 at 24 h with all at 8 h. Conclusion Akt signaling pathway might play an important role in the PASMCs proliferation and differentiation during hypoxia pulmonary hypertension. Aktl may be the most important one.