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中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (3): 218-221.doi: 10.12047/j.cjap.5442.2017.054

• 研究论文 • 上一篇    下一篇

缺氧肺动脉平滑肌细胞中PKCα/c-fos、Bax/Bcl-2表达变化及其对增殖和凋亡的影响

曹艳红1, 潘建勋1, 梁月琴2, 师锐赞2   

  1. 1. 大同市第五人民医院, 山西 大同 037000;
    2. 山西医科大学, 太原 030001
  • 收稿日期:2016-03-28 修回日期:2016-12-07 出版日期:2017-05-28 发布日期:2018-06-20
  • 通讯作者: 师锐赞,Tel:0351-4135079,E-mail:shiruizan@163.com E-mail:shiruizan@163.com
  • 基金资助:
    山西省青年科技研究基金资助项目（2014021038-3）；山西医科大学校科技创新基金资助项目（01201308）；山西医科大学博士启动基金资助项目（B032001207）

Effects of PKCα/c-fos, Bax/Bcl-2 expression on the proliferation and apoptosis of rat pulmonary arterial smooth muscle cells in hypoxia

CAO Yan-hong1, PAN Jian-xun1, LIANG Yue-qin2, SHI Rui-zan2   

  1. 1. Datong the Fifth People's Hospital, Datong 037000;
    2. Shanxi Medical University, Taiyuan 030001, China
  • Received:2016-03-28 Revised:2016-12-07 Online:2017-05-28 Published:2018-06-20
  • Supported by:
    山西省青年科技研究基金资助项目（2014021038-3）；山西医科大学校科技创新基金资助项目（01201308）；山西医科大学博士启动基金资助项目（B032001207）

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摘要： 目的：研究缺氧大鼠肺动脉平滑肌细胞（PASMCs）PKCα/c-fos、Bax/Bcl-2表达的变化及对其增殖和凋亡的影响。方法：将原代培养的大鼠PASMCs分为常氧组（5% CO2、21% O2、74% N2）和低氧组（5% CO2，2% O2，93% N2）24 h，48 h。通过氮蓝四唑盐（MTT）比色法测定PASMCs的增殖水平，以流式细胞仪Annexin V-FITC/PI染色法检测细胞凋亡率，并通过用RT-PCR和Western blot法检测PKCα/c-fos、Bax/Bcl-2基因和蛋白表达的变化。实验至少重复3次，每组设置至少3个复孔。结果：低氧24 h和48 h组PASMCs增殖明显，与常氧组相比差异具有统计学意义（P < 0.01），而细胞凋亡率无显著变化。相对于常氧组，低氧24 h和48 h组PASMCs PKCα、c-fos表达增强，Bax表达变化不明显，而Bcl-2表达增加。结论：低氧培养的PASMCs增殖明显，凋亡无明显变化。同时PKCα、c-fos表达增强，而抗凋亡基因Bcl-2表达增加。

关键词: PKCα/c-fos, Bax/Bcl-2, 增殖, 凋亡, 低氧

Abstract: Objective: To explore the effects of PKCα/c-fos, Bax/Bcl-2 on the proliferation and apoptosis of rat pulmonary arterial smooth musclecells(PASMCs) in hypoxia. Methods: The PASMCs of rats had been isolated and cultured, and then were cultured under normoxia (5%CO2、21%O2、74%N2)and hypoxia(5%CO2,2% O2,93%N2)condition for 24 h and 48 h, respectively. The proliferation of PASMCs was tested by methylthiazolyltetrazolium (MTT). The changes of PASMCs apoptosis were detected by Annexin V-FITC/PI staining combined flow cytometry. RT-PCR and Western blot analysis were performed to detect the gene and protein levels of PKCα/c-fos, Bax/Bcl-2, respectively. All experiments were repeated three times with at least triplicate samples. Results: The proliferation of PASMCs in hypoxia group were stronger than that of normoxia group after 24 h and 48 h (P < 0.01), while the apoptosis rate did not change significantly. Meanwhile, the higher levels of PKCα, c-fos, Bcl-2 mRNA and proteins after hypoxia for 24 h and 48 h were detected by RT-PCR and Western blot analysis, while the ex-pression levels of Bax had no significant difference under normoxic and hypoxic conditions. Conclusion: Elevated proliferation and expression of PKCα, c-fos, Bcl-2 was observed in rat PASMCs in hypoxia, and while the apoptosis rate had no significant change.

Key words: PKCα/c-fos, Bax/Bcl-2, proliferation, apoptosis, hypoxia

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