摘要
目的探讨Fas、FasL在Na+/H+交换器1(NHE1)抑制所诱导的缺氧大鼠肺动脉平滑肌细胞(PASMCs)凋亡中的作用。方法将转染NHE1特异性核酶基因的大鼠PASMCs置于缺氧条件下(O2的体积分数低于1%)培养。缺氧培养2、6、12、24和48h后用原位末端标记法(TUNEL)检测细胞凋亡情况;半定量逆转录聚合酶链反应(sqRTPCR)方法检测细胞内fas和fasLmRNA表达变化;免疫细胞化学法检测细胞内Fas和FasL蛋白表达变化。结果转染NHE1特异性核酶基因的大鼠PASMCs在缺氧培养时,其细胞凋亡率随缺氧时间的延长而逐渐升高,但细胞内fas、fasLmRNA及Fas、FasL蛋白表达与对照组细胞比较差异均无显著性。结论Fas/FasL死亡通路可能不参与NHE1抑制而诱导缺氧大鼠PASMCs凋亡的调控。
Objective To investigate the role of Fas and FasL expression in apoptosis of pulmonary artery smooth muscle cells （PASMCs） in hypoxia induced by inhibition of Na^＋/H^＋ exchanger isoform-1 （NHE-1） in rat. Methods The cells were cultured in hypoxia （〈1% O2） for 2,6,12,24 and 48 hours respectively. Then cell apoptosis was observed with terminal deoxynudeotidyl transferase - mediated dUTP- biotin nick end labeling （TUNEL）. The changes in fas and fasl mRNA expression in PASMCs were detected by semi-quantitative reverse transcription - polymerase chain reaction （sqRT-PCR） in vitro. The expression of Fas and Fasl. protein in cells was determined immunohistochemically. Results No significant changes in the expression of fas/fasL mRNA and protein between cells transfected with NHE - 1 ribozyme gene and cells transfectsd with pLXSN, or nontransfected control in hypoxia. Conclusion Apoptosis of PASMCs induced by NHE - 1 inhibition may occur independently of Fas/FasL death pathway.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2005年第9期515-518,F0005,共5页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目(39870352)
作者简介
陆俊羽（1972-）。女（汉族），云南省人，讲师，医学博士，主治医师，主要从事缺氧肺动脉高雎斤面的研究，已发表论文10篇（Email：lujunyu1026@yahoo．com．cn）。
