低氧肺动脉高压大鼠肺动脉HO-1基因表达和凋亡-第三军医大学学报.DOCVIP

低氧肺动脉高压时血红素氧合酶基因表达状况的研究 王关嵩1，钱 频1Δ，李琦1，李运成1，程序2，周长喜1，张志远1，钱桂生1 (1第三军医大学附属新桥医院全军呼吸内科研究所，重庆 400037；2重庆市垫江县中医院，重庆408300) 摘 　要:目的 探讨血红素氧合酶（heme oxygenase,HO）基因在低氧肺动脉高压大鼠肺动脉组织中的表达及一氧化碳（carbon monoxide，CO）对其的影响。方法 60只 Wistar大鼠随机分为5组：正常对照组(I)；低氧肺动脉高压组(II)；血晶素组(III)；锡原卟啉组(IV)；低浓度一氧化碳（V) 组；每组12只。采用紫外分光光度计、逆转录－聚合酶链反应、免疫组织化学染色和原位杂交进行检测。结果 （1）HO－1活性：低氧组、血晶素组、低浓度CO组肺动脉组织HO-1活性以胆红素生成量计算，均升高，其中血晶素组最高，与正常对照组比较，差异有显著性(P ＜0.01)。（2）逆转录－聚合酶链反应：各组大鼠肺动脉HO－2mRNA表达没有明显变化，均保持稳定。缺氧组、血晶素组、锡原卟啉组和低浓CO组HO－1 mRNA均明显高于正常对照组，以低浓度CO组升高最明显( P ＜0.01)。血晶素组和低浓度CO组均显著高于缺氧组( P ＜0.01)，而锡原卟啉组低于缺氧组。（3）免疫组织化学染色：低氧组肺动脉内膜、中膜的HO-1表达均明显升高，血晶素组和低浓度CO组更高，锡原卟啉组和常氧组均有很微弱表达。（4）原位杂交：低氧组肺动脉三层均加深2-3级,低浓度CO组肺动脉外、中、内膜胞浆均明显加深4级。处理前后HO-2的染色变化不大。结论 低氧肺动脉高压时，肺动脉组织HO－1基因的表达升高，可能在肺动脉高压的发生和发展中起一定作用。 关键词: 血红素氧合酶；低氧；肺动脉高压；基因表达 中图法分类号:R394.2；R543.2 文献标识码:A Study on expression of heme oxygenase in pulmonary arteries of rats with hypoxic pulmonary hypertension WANG Guan-song1, Qian Pin1, LI Qi1, LI Yun-cheng1, CHENG Xu2, ZHOU Chang-xi1, ZHANG Zhi-yuan1, QIAN Gui-sheng1 (1Institute of Respiratory Diseases, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China； 2Chinese Medicine Hospital of Dianjiang County，Chongqing 408300, China) AbstractObjective　To investigate the expression of heme oxygenase (HO) in pulmonary arter of rats with hypoxic pulmonary hypertension. Methods　A total of 60 rats were employed and equally divided into 5 groups, i. e. control(group1, n=12), HPH(group2, n=12), hemin(group3, n=12), Tin Protoporphyrin (Snpp) (group4, n=12) and low concentration CO(group5, n=12) groups. Of them, the rats from middle 3 groups were treated with hypoxia under normal pressure for 7 h in every day except Sunday, 3 weeks. Hemin and Tin Protoporphyrin (Snpp) were given half an hour before hypoxia while low concentration CO after 2 h of hypoxia. HO－1 activity of pulmonary arteries tissues was tested with ultraviolet spectrophotometry. The expression of HO was detected with reverse transcriptionpolymerase chain r