低氧诱导因子-1α双突变体对大鼠缺血心肌VEGF表达及能量代谢的影响

Effects of the recombinant adenovirus containing the 564/402-point mutant HIF-1α gene on VEGF expression and energy metabolism in ischemic myocardium of rats

目的研究低氧诱导因子-1α双突变体(Ad-HIF-1α-564/402)对大鼠缺血心肌血管内皮生长因子(VEGF)的表达及能量代谢的影响。方法构建Ad-HIF-1α-564/402,39只雄性SD大鼠结扎冠状动脉左前降支建立心肌梗死模型,随机分成3组:生理盐水组(Saline组,n=13)、腺病毒空载体组(AdNull组,n=13)、腺病毒HIF-1α双突变组(Ad-HIF-1α-564/402组,n=13)。术后即刻以肌肉注射的方式分别在大鼠缺血心肌给予生理盐水、Ad-Null和Ad-HIF-1α-564/402 0.1 m L,病毒滴度为2.0×108PFU。术后3 d Real-time PCR检测缺血心肌局部VEGF在mRNA水平的表达。术后28 d对比各组急性心肌梗死大鼠基因转染前后体重差、心体比、左心质量与全心质量比;分光光度计法检测大鼠缺血心肌Na+-K+ATP酶、Ca2+-ATP酶、总ATP酶、琥珀酸脱氢酶(SDH)水平;苏木精-伊红(HE)染色法光镜下观察大鼠心肌形态学改变。结果基因转染后3 d,Ad-HIF-1α-564/402组缺血心肌VEGF在mRNA水平表达高于其他两组(P<0.05);基因转染后28 d,3组间体重变化无明显差异(P>0.05),Ad-HIF-1α-564/402组心体比及左心与全心质量比略低于其他两组,差异无统计学意义(P>0.05)。总ATP酶、Na+-K+ATP酶3组间差异无统计学意义(P>0.05),Ad-HIF-1α-564/402组Ca2+-ATP酶、SDH高于其他两组(P<0.05)。心肌组织HE染色未见明显肿瘤样组织生成。结论 Ad-HIF-1α-564/402能上调缺血心肌VEGF在mRNA水平的表达,改善能量代谢,有一定的安全性。

Objective To investigate the effects of the recombinant adenovirus containing the 564/402-point mutant HIF-1α gene（ Ad-HIF-1α-564/402） on VEGF expression and energy metabolism in ischemic myocardium of rats. Methods Ad-HIF-1α-564/402 was constructed. The model of myocardial infarction was established by ligation of the left anterior descending coronary artery in thirty-nine male Sprague-Dawley rats. The rats were randomly divided into 3 groups,Saline group（ n = 13）,Ad-Null group（ n = 13）,and Ad-HIF-1α-564/402 group（ n = 13）.The ischemic myocardium received intramuscular injection of saline,Ad-Null,or Ad-HIF-1 ar mi/402 0. 1 m L（ 1. 0 ar8 PFU）,respectively,in the 3 groups. Three days after gene transfection,real-time PCR was used to assess the RNA expression of VEGF in the ischemic myocardium. Twenty-eight days after gene transfection,the differences in body weight,body mass ratio,and left ventricular mass to total heart mass were compared. The Na＋-K＋ATPase,Ca2 ＋-ATPase,total ATPase,and Succinate dehydrogenase（ SDH） levels were detected by spectrophotometer. The morphological changes of myocardium were observed under light microscope with hematoxylin-eosin（ HE） staining. Results On the3 thday after gene transfection,the mRNA expression of VEGF in Ad-HIF-1α-564/402 group was significantly higher than the other two groups（ P〈0. 05）. On the 28 thday after gene transfection,there was no significant difference in the weight among the three groups. Body mass ratio and left ventricular mass to total heart mass om HIF-1α-564/402 group were slightly lower than the other two groups,but the difference was not statistically significant. There was no significant difference in total ATPase or Na＋-K＋ATPase among the three groups. The Ca^2 ＋-ATPase and SDH in Ad-HIF-1α-564/402 group were significantly higher than the other two groups（ P〈0. 05）. The hematoxylin-eosin（ HE） staining in myocardial tissue showed no obvious tumor tissue. Conclusion Ad-HIF-1α-564/402 can upregulate the expression of VEGF in ischemic myocardium,and improve energy metabolism,with certain safety.