eNOS基因转染对慢性缺氧性肺动脉高压的影响

Effect of adenovirus-mediated human endothelial nitric oxide synthase gene transfer on chronic hypoxic pulmonary hypertension

目的:观察重组缺陷性腺病毒介导的人内皮型一氧化氮合酶基因(AdCMVceNOS)对慢性缺氧性肺动脉高压大鼠的影响,探讨其可能的作用机制.方法:雄性W istar大鼠30只,随机分为对照组(C组)、单纯慢性低氧组(H组)、慢性低氧+AdCMVceNOS转染组(T组),每组10只.采用呼吸道转染途径,将3×109空斑形成单位的AdCMVceNOS转入大鼠肺脏.3 d后测平均肺动脉压、右心室重量及肺动脉血NO含量,并采用免疫组化及W estern b lot方法观察内皮型一氧化氮合酶(eNOS)的表达情况.结果:H组及T组平均肺动脉压(mPAP)均较C组明显增加(P<0.05);T组与H组比较mPAP降低(P<0.05);H组及T组右心室重量/(左心室+室间隔)重量较C组增加(P<0.05).H组与C组相比,其NO的含量明显降低(P<0.05),T组较H组显著增加(P<0.05).H组及T组eNOS阳性表达强度(A值)较C组明显增加(P<0.05),而T组与H组相比差异有统计学意义(P<0.05).W estern b lot检测H组及T组肺组织eNOS含量分别为0.62±0.08和0.92±0.17,明显高于C组0.30±0.05.结论:外源性eNOS基因转染对大鼠慢性缺氧性肺动脉高压具有治疗作用,该作用与其使eNOS活性增强及NO生成增加有关.

AIM： To investigate the effect of adenovirusmediated human endothelial nitric oxide synthase （eNOS） gene （AdCMVceNOS） transfered to the rat lungs through airway on chronic hypoxic pulmonary hypertension （CHPH） rats and to explore its possible role in CHPH therapy. METHODS： Thirty male Wistar rats were randomly divided into 3 groups（n = 10 per group）, the control group（ group C）, the chronic hypoxic group （group H）, the chronic hypexic and AdCMVceNOS transferred group（group T）. The AdCMVceNOS （3 -10^9 pfu） were tranfeted into the rat lungs through the respiratory tract. Survey the pulmonary artery pressure （ PAP ） , right ventricular weight and content of NO in pulmonary artery respectively after 3 d. The expression activity of eNOS was observed by immunohistochemistry and Western blot methods. RESULTS： The mean PAP and the RV/（LV ＋ S） of group H and T increased obviously compared with group C （ P 〈 0. 05 ） ; The mean PAP of group T decreased compared with group H （ P 〈 0.05 ） ; Compared with group C, the content of NO in group H decreased obviously （ P 〈 0.05 ）, but group T was increased more obviously than group H （ P 〈 0.05 ）. The A value of eNOS in group H and T increased obviously compared with that of group C （ P 〈 0.05 ）, but the group T was significantly different from group H in A value （ P 〈 0.05 ）. The levels of eNOS protein in group H （0.62 3：0.08 ） and T （ 0. 92 3： 0.17）were much higher than that in group C （0.30±0.05）. CONCLUSION： Gene transfer of eNOS to the lung may be a useful therapeutic intervention for the treatment of CHPH, which may be correlated with the increased eNOS activity and NO content.