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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-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 heme oxygenase (HO) gene in pulmonary artery of hypoxic pulmonary hypertensive rats and the effect of carbon monoxide (CO) on it. Methods Sixty Wistar rats were randomly divided into 5 groups: normoxia group, hypoxic pulmonary hypertension group, hemin group, tin protoporphyrin group and low concentration CO group (n = 12). UV spectrophotometer, reverse transcription - polymerase chain reaction, immunohistochemical staining and in situ hybridization were used for detection. Results The activity of HO-1 in HO-1 pulmonary artery tissue of hypoxia group, hemin group and low-concentration CO group were all increased with the increase of bilirubin production, the highest in hemin group. Compared with normal control group, The difference was significant (P <0.01). Reverse transcriptase-polymerase chain reaction: There was no significant change in the expression of HO-2 mRNA in pulmonary arteries of rats in each group, both of which remained stable. HO-1mRNA in hypoxia group, hemin group, protoporphyrin group and low-concentration CO group were significantly higher than those in normal control group, and the most obvious increase was in low-concentration CO group (P <0.01). Hemin group and low concentration CO group were significantly higher than hypoxia group (P <0.01), while tin protoporphyrin group was lower than hypoxia group. ③ Immunohistochemical staining: The expression of HO-1 in pulmonary arterial intima and media in hypoxia group was significantly higher than that in hypochlorocytoma group and low-concentration CO group, both in protoporphyrin group and normoxia group expression. ④ In-situ hybridization: The pulmonary arteries in hypoxia group were all deepened 2-3 levels. The pulmonary cytoplasm outside, middle and intima of CO group were significantly deepened 4 levels in low concentration CO group. The staining of HO-2 did not change much before and after treatment. Conclusions HO-1 gene expression in pulmonary artery is increased in hypoxic pulmonary hypertension, which may play a role in the occurrence and development of pulmonary hypertension.