低氧肺动脉高压时血红素氧合酶基因表达状况的研究  

Expression of heme oxygenase in pulmonary arteries of rats with hypoxic pulmonary hypertension

作　　者：王关嵩[1] 钱频[1] 李琦[1] 李运成[1] 程序 周长喜[1] 张志远[1] 钱桂生[1] 

机构地区：[1]第三军医大学新桥医院全军呼吸内科研究所,全军呼吸病研究重点实验室,重庆400037 [2]重庆市垫江县中医院呼吸内科,重庆408300

出　　处：《第三军医大学学报》2009年第15期1428-1431,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30770928);重庆市自然科学基金(2007BB5072);第三军医大学科研创新基金(2006HG04)~~

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：常氧组；低氧肺动脉高压组；血晶素组；锡原卟啉组；低浓度CO组（n=12）。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①HO-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-1mRNA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3层均加深2～3级，低浓度CO组肺动脉外、中、内膜细胞质均明显加深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。Objective To investigate the expression of heine oxygenase （HO） in pulmonary arteries of rats with hypoxic pulmonary hypertension （HPH）. Methods A total of 60 rats were employed and equally divided into 5 groups （n =12 in each group）, i. e. control （group Ⅰ ）, HPH （group Ⅱ ）, hemin （group Ⅲ）, tin protoporphyrin （SnPP） （group Ⅳ）, and low concentration CO group （group Ⅴ ）. Of them, the rats from middle 3 groups were treated with hypoxia under normal pressure for 7 h in every day except Sunday, consecutively for 3 weeks. Heroin and tin protoporphyrin （SnPP） were given half an hour before hypoxia while low concentration CO at 2 h after hypoxia. HO-1 activity of pulmonary arteries tissues was tested with ultraviolet spectrophotometry. The expression of HO was detected with reverse transcriptionpolymerase chain reaction （RT-PCR） , immunohistochemical staining and in situ hybridization. Results ① The bilirubin production increased significantly in group [ （396.28 ± 69.62） pmol . mg^-1 . h^-1], group [ （668.56 ± 189.29 ） pmol . mg^-1 . h^-1], group [ （267.38±88.59） pmol . mg^-1 . h^-1 ] and group [ （589.32 ± 132.35） pmol . mg^-1.h^-1 ], as com- pared with those in group [ （125.62 ± 31.52） pmol . mg^-1 . h^-1 ]. ② The levels of HO-1 mRNA expression of pulmonary arteries in group Ⅱ , group Ⅲ, group Ⅳ and group Ⅴ were significantly higher than those in group Ⅰ .The most significant HO-1 mRNA expression was found in group Ⅴ （P 〈0.01 ）. The levels of HO-1 mRNA expression of pulmonary arteries in group Ⅲ and group V were higher than those in group Ⅱ , but it was lower in group Ⅳ than that in groupⅡ. ③ The strong positive granules of HO-1 were observed in pulmonary arteries intima and media in group Ⅱ , group Ⅲ and group Ⅳ, while group m and group Ⅳ involved more positive granules of HO-1 as compared with those in group Ⅱ. ④ The staining was deepened in three layers of the pulmonary arteries in group Ⅱ ,

关 键 词：血红素氧合酶 低氧 肺动脉高压 基因表达 

分 类 号：R345[医药卫生—基础医学] R363.25