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目的观察腺病毒介导的内皮型一氧化氮合酶基因(eNOS)转染对高肺血流肺动脉高压大鼠模型肺动脉高压形成的影响。方法随机将40只大鼠分为假手术组、手术组、实验组和对照组,每组10只。手术组、实验组和对照组采用套管连接法建立左向右分流肺动脉高压模型,实验组采用气管吸入法转染重组腺病毒AdCMVeNOS,采用免疫组化法观察eNOS蛋白表达,采用比色法测定肺组织eNOS活性,测量各组大鼠平均肺动脉压(mPAP)、右心室平均压(mRVP)、平均颈动脉压(mCAP)、右心肥厚指数(RVHI),采用HE染色观察肺动脉形态学改变,计算管壁厚度与血管外径比值(WT%)。结果实验组免疫组化染色可见全层血管壁内棕黄色颗粒,eNOS表达强度明显高于对照组(P<0.05),假手术组eNOS表达的阳性细胞介于实验组与对照组之间。手术组和对照组的eNOS活性比假手术组明显降低(P<0.05);实验组的eNOS活性显著高于对照组,且较假手术组亦明显升高(P<0.05)。手术组和对照组大鼠的mRVP、mPAP、RVHI和WT%均明显高于假手术组(P<0.05)。与对照组相比,实验组大鼠mRVP、mPAP、RVHI和WT%均显著降低(P<0.05);手术组与对照组各值之间差异不明显。结论腺病毒介导的eNOS基因转染能够上调高肺血流肺动脉高压大鼠肺组织中eNOS表达及其活性,延缓肺动脉高压的发展和肺血管的重塑。
Objective To investigate the effect of adenovirus-mediated endothelial nitric oxide synthase (eNOS) transfection on pulmonary hypertension in a rat model of pulmonary hypertension induced by high pulmonary blood flow. Methods Forty rats were randomly divided into sham operation group, operation group, experimental group and control group, with 10 rats in each group. The left and right shunt pulmonary hypertension models were established in the operation group, experimental group and control group by using the cannula connection method. The experimental group was transfused with recombinant adenovirus AdCMVeNOS by tracheal aspiration method and the expression of eNOS protein was observed by immunohistochemical method. The eNOS activity in lung tissue was measured. The mean pulmonary arterial pressure (mPAP), mRVP, mCAP and RVHI in each group were measured. Morphological changes of pulmonary arteries were observed by HE staining. The ratio of wall thickness to vessel diameter was calculated (WT%). Results The expression of eNOS in the whole blood vessel wall of the experimental group was observed by immunohistochemistry. The eNOS expression intensity of the experimental group was significantly higher than that of the control group (P <0.05). The positive cells of eNOS expression in the sham operation group were between the experimental group and the control group. The eNOS activity in the surgery group and the control group was significantly lower than that in the sham operation group (P <0.05). The eNOS activity in the experimental group was significantly higher than that in the sham operation group and also significantly higher than that in the sham operation group (P <0.05). The mRVP, mPAP, RVHI and WT% of the rats in the surgery group and the control group were significantly higher than those in the sham operation group (P <0.05). Compared with the control group, the mRVP, mPAP, RVHI and WT% in the experimental group were significantly decreased (P <0.05); there was no significant difference between the operation group and the control group. Conclusion Adenovirus-mediated eNOS gene transfection can up-regulate the expression and activity of eNOS in lung tissue of pulmonary artery hypertension rats and delay the development of pulmonary hypertension and pulmonary vascular remodeling.