Cao Wen, Chang Ya-Fei, Zhao Ai-Chao, Chen Bang-Dang, Liu Fen, Ma Yi-Tong, Ma Xiang
Department of Cardiology, Second People's Hospital of Wuxi, 68 Zhongshan Road, Wuxi, 214000, China.
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, 137 Liyushan Road, Urumqi, 830054, China.
J Mol Histol. 2017 Aug;48(4):275-283. doi: 10.1007/s10735-017-9725-4. Epub 2017 Jun 22.
The present study aimed to investigate the protective effects of rAAV9-CyclinA2 combined with fibrin glue (FG) in vivo in rats after myocardial infarction (MI). Ninety male Sprague-Dawley rats were randomized into 6 groups (15 in each group): sham, MI, rAAV9-green fluorescent protein (GFP) + MI, rAAV9-CyclinA2 + MI, FG + MI, and rAAV9-CyclinA2 + FG + MI. Packed virus (5 × 10vg/ml) in 150 µl of normal saline or FG was injected into the infarcted myocardium at five locations in rAAV9-GFP + MI, rAAV9-CyclinA2 + MI, and rAAV9-CyclinA2 + FG + MI groups. The sham, MI, and FG + MI groups were injected with an equal volume of normal saline or FG at the same sites. Five weeks after injection, echocardiography was performed to evaluate the left ventricular function. The expressions of CyclinA2, proliferating cell nuclear antigen (PCNA), and phospho-histone-H3 (H3P), vascular density, and infarct area were assessed by Western blot, immunohistochemistry, immunofluorescence, and Masson staining. As a result, the combination of rAAV9-CyclinA2 and FG increased ejection fraction and fractional shortening compared with FG or rAAV9-CyclinA2 alone. The expression level of CyclinA2 was significantly higher in the rAAV9-CyclinA2 + FG + MI group compared with the rAAV9-CyclinA2 + MI and FG + MI groups (70.1 ± 1.86% vs. 14.74 ± 2.02%, P < 0.01; or vs. 50.13 ± 3.80%; P < 0.01). A higher expression level of PCNA and H3P was found in the rAAV9-CyclinA2 + FG + MI group compared with other groups. Comparing with other experiment groups, collagen deposition and the infarct size significantly decreased in rAAV9-CyclinA2 + Fibrin + MI group. The vascular density was much higher in the rAAV9-CyclinA2 + FG + MI group compared with the rAAV9-CyclinA2 + MI group. We concluded that fibrin glue combined with rAAV9-CyclinA2 was found to be effective in cardiac remodeling and improving myocardial protection.
本研究旨在探讨重组腺相关病毒9型-细胞周期蛋白A2（rAAV9-CyclinA2）联合纤维蛋白胶（FG）在大鼠心肌梗死（MI）后体内的保护作用。90只雄性Sprague-Dawley大鼠被随机分为6组（每组15只）：假手术组、MI组、rAAV9-绿色荧光蛋白（GFP）+MI组、rAAV9-CyclinA2+MI组、FG+MI组和rAAV9-CyclinA2+FG+MI组。在rAAV9-GFP+MI组、rAAV9-CyclinA2+MI组和rAAV9-CyclinA2+FG+MI组的梗死心肌五个部位注射150μl生理盐水中的浓缩病毒（5×10vg/ml）或FG。假手术组、MI组和FG+MI组在相同部位注射等体积的生理盐水或FG。注射后5周，进行超声心动图检查以评估左心室功能。通过蛋白质免疫印迹法、免疫组织化学、免疫荧光和Masson染色评估细胞周期蛋白A2、增殖细胞核抗原（PCNA）和磷酸化组蛋白H3（H3P）的表达、血管密度和梗死面积。结果显示，与单独使用FG或rAAV9-CyclinA2相比，rAAV9-CyclinA2与FG联合使用可提高射血分数和缩短分数。与rAAV9-CyclinA2+MI组和FG+MI组相比，rAAV9-CyclinA2+FG+MI组中细胞周期蛋白A2的表达水平显著更高（70.1±1.86%对14.74±2.02%，P<0.01；或对50.13±3.80%；P<0.01）。与其他组相比，rAAV9-CyclinA2+FG+MI组中PCNA和H3P的表达水平更高。与其他实验组相比，rAAV9-CyclinA2+纤维蛋白+MI组的胶原沉积和梗死面积显著减少。与rAAV9-CyclinA2+MI组相比，rAAV9-CyclinA2+FG+MI组的血管密度更高。我们得出结论，纤维蛋白胶联合rAAV9-CyclinA2在心脏重塑和改善心肌保护方面是有效的。
