腺病毒介导的鞘氨醇激酶基因转染对心肌梗死后心衰的改善作用研究

Effect of transfection of adenovirus-mediated sphingosine kinase gene on amelioration of post-infarction heart failure

目的探索腺病毒介导的鞘氨醇激酶1(SPK1)基因局部表达对心肌梗死后心衰的治疗作用。方法将20只Wistar大鼠(250～300g)随机分成3组:假手术组(6只)、Ad-GFP对照组(7只)和Ad-SPK1(7只)组。结扎Wistar大鼠冠状动脉左前降支,Ad-SPK1组在心脏梗死区及周围多位点注射携带人SPK1基因的复制缺陷型重组腺病毒(Ad-SPK1),Ad-GFP对照组注射同体积的携带绿色荧光蛋白(GFP)基因的重组腺病毒(Ad-GFP),14d后进行血流动力学及组织形态学检查。结果 Ad-SPK1组左心室收缩压(LVSP)、左心室舒张末压(LVEDP)、最大左心室收缩压上升/降低时间(±dP/dtmax)(分别为132.82±13.03mmHg、4.34±0.69mmHg、5095.20±384.79)与Ad-GFP组(分别为76.96±8.44mmHg、14.79±1.08mmHg、2954.12±195.05)比较明显改善(P<0.01),而与假手术组(分别为147.42±10.92mmHg、2.80±0.45mmHg、5865.19±484.36)比较则无明显差异。Ad-SPK1组梗死区面积(3.78%±0.96%)与Ad-GFP组(38.86%±5.68%)比较明显减小(P<0.01)。Ad-SPK1组左心室直径、左心室壁厚度(分别为4.63±0.80mm、2.70±0.29mm)与Ad-GFP组(分别为7.30±1.03mm、1.34±0.36mm)比较,直径明显缩小,厚度变薄的程度减轻(P<0.01),而与假手术组(分别为4.50±0.36mm、2.80±0.34mm)比较则无明显差异。Ⅷ因子相关抗原免疫组化分析显示,SPK1基因转染能明显刺激梗死缺血区的血管生成;天狼星红染色结果表明,SPK1基因转染能显著降低胶原在梗死缺血区的沉积。结论腺病毒介导的SPK1基因转染能够保护缺血导致的心功能损伤,局部Ad-SPK1注射可能是治疗缺血性心脏病的一条新途径。

Objective To investigate the therapeutic effect of adenovirus-mediated sphingosine kinase 1gene（Ad-SPK1）transfection on amelioration of post-infarction heart failure.Methods Twenty Wistar rats（250-300g）were randomized to three groups：sham group （n=6）,adenovirus-mediated green fluorescent protein group（Ad-GFP,n=7）and Ad-SPK1group（n=7）.The left anterior descending branch of coronary artery was ligated,followed by direct intramyocardial injection of AD-SPK1or Ad-GFP gene in multiple sites as control.The hemodynamics and histomorphology of heart was analyzed 14days after operation.Results Left ventricular systolic pressure （LVSP）,left ventricular end-diastolic pressure（LVEDP）and maximum dP/dt in Ad-SPK1group（132.82±13.03mmHg,4.34± 0.69mmHg and 5095.20±384.79,respectively）were significantly improved compared with that in Ad-GFP group（76.96±8.44mmHg, 14.79±1.08mmHg and 2954.12±195.05,respectively,P〈0.01）,but there were no significant differences between Ad-SPK1group and sham group（147.42±10.92mmHg,2.80±0.45mmHg and 5865.19±484.36,respectively）.Infarct size in Ad-SPK1group（3.78%± 0.96%）was significantly smaller than that in Ad-GFP group（38.86%±5.68%,P〈0.01）.Left ventricular（LV）diameter and LV wall thickness in Ad-SPK1group（4.63±0.80mm,2.70±0.29mm,respectively）were significantly smaller than that in Ad-GFP group （7.30±1.03mm,1.34±0.36mm,respectively,P〈0.01）,but no differences were observed between Ad-SPK1group and sham group （4.50±0.36mm,2.80±0.34mm）.Transfection of SPK1gene enhanced angiogenesis significantly as revealed by Von Willebrand’s factor immunohistochemical staining and blood vessel counting,and reduced fibrosis shown by Sirius red staining.Conclusions Transfection of adenovirus-mediated SPK1gene may efficiently improve postischemic heart failure by enhancing angiogenesis and reducing fibrosis, implying that the transfection of adenovirus-mediated SPK1gene might provide a novel strategy for treatment of coronary heart disease.