摘要
目的探讨重组腺病毒介导的人组织激肽释放酶(h TK1)基因过表达对大鼠心肌梗死后心肌结构、心肌胶原纤维以及新生血管的影响。方法结扎雄性Sprague-Dawley大鼠的左冠状动脉前降支,建立急性心肌梗死模型。将含h TK1基因的重组腺病毒(Ad-h TK1)和对照病毒(1×1010PFU)注射到心肌梗死部位及其周边区域。4周后处死大鼠,取出心脏、称量、切片;通过荧光显微镜观察心肌组织的绿色荧光表达;Western blot测定h TK1蛋白表达;HE染色观察心肌组织结构,天然猩红染色观察胶原含量、分布,免疫组化法检测新生血管密度。结果仅在Ad-h TK1组大鼠的心肌既有绿色荧光,又有h TK1蛋白表达。假手术组大鼠的心脏质量、左室质量、心肌胶原分布显著低于心梗模型组,新生血管密度明显高于模型组。但与对照病毒组相比,Ad-h TK1组大鼠的心脏质量、左室质量、左室质量指数和心肌胶原纤维含量明显降低(P<0.05),心肌结构和新生血管密度明显改善(P<0.05)。结论重组腺病毒介导的组织激肽释放酶基因过表达可改善心肌结构,增加梗死周围心肌的新生血管形成,改善梗死后心肌重构。
Objective To explore the effect of recombinant adenovirus-mediated human tissue kallikrein- 1 （Ad-hTK1） on ventricular remodeling in rats after myocardial infarction （MI）. Methods Rats were subjected to ligating left anterior descending artery of coronary artery. 1 × 10^10 PFU Ad- hTK1 or control virus （Ad-EGFP） were administrated at multiple sites into the infarcted myocardium 1 h after the operation. Four weeks after the in- tervention, the protein expression of hTK1 was evaluated by Western blot analysis. Expression of the EGFP in MI was detected by fluorescence mi- croscopy. Histological morphometric observation was carried out using fluorescence microscope and HE staining. Collagen were determined by Sirius red-saturated picric staining, and the number of myocardial microvessel was detected by CD34-FITC antibody immunocytochemistry assay. Results The expression of green fluorescenee and hTK1 protein were observed in Ad-hTK1 rats. There were no differences in body weight among different groups at 4 weeks after MI. As compared with sham group, MI resulted in increases in heart weight, LVW and LVWI, inflammation cells and colla- gen density, in decreases capillary density, at 4 weeks after MI. However, hTK1 gene delivery tended to reduce heart weight, LVW, LYWI, inflam- marion cells and collagen density at 4 weeks after MI. Capillary density in MI- hTK1 rots was also significantly increased than in the M1 with control virus. Conclusion This study indicates that recombinant adenovirus-mediated human tissue kallikrein- 1 overexpression plays an important role in preventing the progression of MI by attenuating cardiac hypertrophy and fibrosis, and improving neovaseularization.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2014年第9期830-835,共6页
Journal of China Medical University
基金
福建省自然科学基金(2011J01134
2010J01127)
福建省卫生厅青年骨干培养人才项目(2013-ZQN-ZD-2)
关键词
人组织激肽释放酶1
重组腺病毒
心肌梗死
心室重构
human tissue kallikrein 1
adenovirus
myocardial infarction
ventricular remodeling
