Li Chi-guan, Dai Ai-guo, Huang Cui-ping
Department of Respiratory Medicine, Hunan Geriatric Hospital, Changsha 410001, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2005 May;21(2):187-91.
To investigate the effects of hypoxia on the proliferation and apoptosis of PASMC, to evaluate the role of iNOS protein expression and ADM on the hypoxic pulmonary hypertension (HPH) pathogenesis.
To culture rat pulmonary artery smooth muscle cell (PASMC), cultured PASMC cells were grouped into: normoxic group; hypoxic group; hypoxia + L-NAME group; hypoxia+ ADM group. Proliferation of PASMC were investigated by MTT and PCNA. Apoptosis of PASMC were examined by flow-cytometry. Westen blot was used to measure protein expression of iNOS induced by hypoxia.
(By MTT, the value of 24 h hypoxia was significantly higher than that in the normoxic group (P < 0.01), the value of the hypoxia + ADM was significantly lower than that in hypoxia group, the value of the hypoxia + L-NAME was significantly higher than those of hypoxic group and normoxic group (P < 0.01). (2) By immunohistochemistry, PCNA was poorly positive in PASMC, whereas positive after 24 h hypoxia (P < 0.01), ADM inhibited the expression of PCNA significantly (P < 0.01), whereas L-NAME increased the expression of PCNA significantly (P < 0.01). (3) By FCM, apoptosis index was not significantly different between the normoxic group, hypoxic group, hypoxia + L-NAME and hypoxia + ADM (P > 0.05). (4) By Western blot, iNOS expression was poorly positive in control group, positive after 4 h hypoxia (P < 0.01), increasing as the hypoxia environment continued (P < 0.01). L-NAME had no effect on iNOS protein, ADM promoted iNOS expression (P < 0.01).
(1) Hypoxia stimulates the proliferation of PASMC, and has no obvious effects on the apoptosis of PASMC. (2) Hypoxia induces the expression of iNOS, ADM can increase expression of iNOS, ADM and INOS plays a role of protection in HPH pathogenesis.
研究缺氧对肺动脉平滑肌细胞（PASMC）增殖和凋亡的影响，评估诱导型一氧化氮合酶（iNOS）蛋白表达及肾上腺髓质素（ADM）在低氧性肺动脉高压（HPH）发病机制中的作用。
培养大鼠肺动脉平滑肌细胞（PASMC），将培养的PASMC细胞分为：常氧组；缺氧组；缺氧+L-硝基精氨酸甲酯（L-NAME）组；缺氧+ADM组。采用噻唑蓝（MTT）法和增殖细胞核抗原（PCNA）检测PASMC的增殖情况。采用流式细胞术检测PASMC的凋亡情况。采用蛋白质免疫印迹法检测缺氧诱导的iNOS蛋白表达。
（1）MTT法检测结果显示，缺氧24小时组的值显著高于常氧组（P<0.01），缺氧+ADM组的值显著低于缺氧组，缺氧+L-NAME组的值显著高于缺氧组和常氧组（P<0.01）。（2）免疫组织化学检测结果显示，PASMC中PCNA弱阳性，缺氧24小时后阳性（P<0.01），ADM显著抑制PCNA的表达（P<0.01），而L-NAME显著增加PCNA的表达（P<0.01）。（3）流式细胞术检测结果显示，常氧组、缺氧组、缺氧+L-NAME组和缺氧+ADM组的凋亡指数差异无统计学意义（P>0.05）。（4）蛋白质免疫印迹法检测结果显示，对照组iNOS表达弱阳性，缺氧4小时后阳性（P<0.01），随着缺氧环境持续表达增加（P<0.01）。L-NAME对iNOS蛋白无影响，ADM促进iNOS表达（P<0.01）。
（1）缺氧刺激PASMC增殖，对PASMC凋亡无明显影响。（2）缺氧诱导iNOS表达，ADM可增加iNOS表达，ADM和iNOS在HPH发病机制中起保护作用。
