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目的:观察重组腺相关病毒介导血红素加氧酶1(AAV-rHO-1)基因在大鼠心肌的表达情况,及其对大鼠心肌离体缺血再灌注损伤的影响。方法:雄性SD大鼠30只随机分成3组,对照组(C组,n=6),缺血再灌注组(I/R组,n=12),AAV-rHO-1基因组(A-r组,n=12)。基因转染3个月后,半定量RT-PCR检测转染基因的表达情况,并建立大鼠离体心脏Langendorff灌流模型,C组持续灌注100 min,其他各组均平衡15 min,停灌40 min与再灌注45 min,记录各组心功能变化,测定冠脉流出液乳酸脱氢酶(LDH)活性,测定再灌注后45 min时的心肌超氧化物岐化酶(SOD)活性及丙二醛(MDA)含量,同时取每组大鼠心肌检测梗死面积。结果:经半定量RT-PCR分析显示,血红素加氧酶在A-r组心肌中有效表达。离体心肌Langendorff灌注时,与I/R组比较,A-r组在复灌后各时间点心功能明显增强(P<0.01),复灌后冠脉流出液LDH活性降低(P<0.01),复灌后45 min后心肌MDA含量降低(P<0.01),SOD活性增高(P<0.01),梗死面积较小(P<0.01)。结论:腺相关病毒携带的血红素加氧酶1基因能有效地转染心肌组织,并在体内稳定表达,AAV-rHO-1转染预处理对大鼠离体心肌缺血再灌注损伤有显著保护作用。
OBJECTIVE: To observe the expression of recombinant adeno-associated virus-mediated heme oxygenase-1 (AAV-rHO-1) gene in rat myocardium and its effect on myocardial ischemia-reperfusion injury in vitro. Methods: Thirty male SD rats were randomly divided into 3 groups: control group (C group, n = 6), ischemia reperfusion group (I / R group, n = 12), AAV-rHO- n = 12). Three months after gene transfection, the expression of transfected gene was detected by semi-quantitative RT-PCR. Langendorff perfusion model was established in isolated rat heart. C group was continuously perfused for 100 min. All the other groups were balanced for 15 min. min and reperfusion for 45 min, the change of cardiac function in each group was recorded. The coronary effluent lactate dehydrogenase (LDH) activity was measured. The activity of myocardial superoxide dismutase (SOD) Aldehyde (MDA) content, while taking the myocardial infarction area of each rat. Results: Semi-quantitative RT-PCR analysis showed that heme oxygenase was efficiently expressed in myocardium of A-r group. Compared with the I / R group, the cardiac function of Ar group was significantly increased at Langendorff perfusion (P <0.01), and the LDH activity of coronary effluent decreased after reperfusion (P <0.01) After 45 min, MDA content decreased (P <0.01), SOD activity increased (P <0.01) and infarct size smaller (P <0.01). CONCLUSION: Adeno-associated virus-carrying heme oxygenase-1 gene can be efficiently transfected into myocardium and stably expressed in vivo. Pretreatment with AAV-rHO-1 transfection can significantly inhibit myocardial ischemia-reperfusion injury in isolated rat hearts Protective effects.