抑制磷酸二酯酶5的短发夹RNA重组腺病毒载体对阿霉素所致心肌病小鼠的保护作用

发布时间：2018-05-29 03:01

本文选题：阿霉素 + 心肌病　；参考：《中国临床药理学杂志》2017年14期

【摘要】：目的研究靶向磷酸二酯酶5(PDE5)抑制剂短发夹RNA(shRNA)[PDE5shRNA]重组腺病毒载体对阿霉素所致心肌病的保护作用。方法用一次性腹腔内注射阿霉素15 mg·kg~(-1)诱导小鼠阿霉素心肌病模型。按照体重将小鼠随机分为3组:模型组(n=16);实验组(阿霉素+PDE5shRNA,n=16)在造模后,一次性心肌多部位注射PDE5shRNA重组腺病毒载体(1×1010粒子/鼠);正常组(n=10):一次性腹腔注射等量0.9%NaCl。给药2周后,对存活小鼠进行心脏超声检查,检测左心室收缩末期内径(LVESD)、左心室舒张末期内径(LVEDD)、左心室射血分数(LVEF)和短轴缩短率(%FS)。以苏木精-伊红(HE)染色法测量心肌细胞直径;以Masson氏染色法测量心肌纤维化面积;以酶联免疫吸附法检测各组小鼠心肌组织中环磷酸鸟苷(c GMP)和蛋白激酶(PKG)含量,以免疫蛋白印迹法检测各组小鼠心肌组织中肌凝蛋白重链(MHC)、肌钙蛋白I(Tropnin1)以及肌间蛋白(Desmin)的表达情况。结果给药后2周后,实验组与模型组的LVESD分别是(2.13±0.10),(2.75±0.12)mm,这2组的LVEDD分别是(2.98±0.10),(3.42±0.12)mm,这2组的LVEF分别是(58.74±1.40)%,(48.53±1.50)%,这2组的%FS分别为(28.52±2.10)%,(19.59±1.67)%,2组比较差异均有统计学意义(均P0.05)。这2组的心肌细胞直径分别是(14.68±0.42),(13.75±0.38)μm,差异有统计学意义(P0.05)。实验组与模型组的心肌纤维化面积比率分别为(2.28±0.20)%,(5.10±0.35)%,差异有统计学意义(P0.05)。这2组的c GMP含量分别是(0.23±0.02),(0.06±0.01)pg·mg~(-1),这2组的PKG含量分别是(0.21±0.02),(0.10±0.01)pg·mg~(-1),2组比较差异均有统计学意义(均P0.05)。实验组与模型组的MHC表达分别是1.55±0.16,1.15±0.22,这2组的Tropnin1表达分别是1.32±0.08,0.88±0.08,这2组的Desmin表达分别是1.33±0.51,1.10±0.04,差异均有统计学意义(均P0.05)。结论 PDE5shRNA可能通过激活c GMP/PKG通路减轻阿霉素所致心肌细胞萎缩和心肌纤维化,明显改善左心室功能障碍。
[Abstract]:Objective to study the protective effect of recombinant adenovirus vector (PDE5shRNA) targeting phosphodiesterase 5 (PDE5) inhibitor, short hairpin RNA-shRNA, on adriamycin-induced cardiomyopathy. Methods the model of adriamycin cardiomyopathy in mice was induced by intraperitoneal injection of adriamycin 15 mg 路kg ~ (-1). The mice were randomly divided into three groups according to their body weight: model group (n = 16), experimental group (adriamycin PDE5 shRNAnn16), single myocardial injection of PDE5shRNA recombinant adenovirus vector 1 脳 1010 seeds / mouse after model making, and normal group (n = 10): intraperitoneal injection of 0.9% NaCl. Two weeks after administration, the left ventricular end-systolic diameter (LVESD), left ventricular end-diastolic diameter (LVEDDD), left ventricular ejection fraction (LVEF) and short axis shortening rate were detected by echocardiography. The diameter of cardiac myocytes was measured by hematoxylin-eosin (HEH) staining, the area of myocardial fibrosis was measured by Masson's staining, and the contents of cyclic guanosine phosphate (CGPP) and protein kinase (PKG) were detected by enzyme-linked immunosorbent assay (Elisa). The expression of myosin heavy chain (MHCC), troponin Itropnin1 (Itropnin1) and intermuscular protein (Desmin) in myocardium of each group was detected by Western blot. Results two weeks after administration, the LVESD of the experimental group and the model group were 2.13 卤0.10 卤0.12 卤2.75 卤0.12 mm, respectively, and the LVEDD of the two groups were 2.98 卤0.10 卤0.12 卤3.42 卤0.12, respectively. The LVEF of the two groups were 58.74 卤1.40 卤48.53 卤1.50, respectively. The FS of the two groups were 28.52 卤2.1010 and 19.59 卤1.67 respectively (P 0.05). The diameter of cardiomyocytes in these two groups was 14.68 卤0.42 卤0.38 渭 m, respectively, and the difference was statistically significant (P 0.05). The areas of myocardial fibrosis in the experimental group and the model group were 2.28 卤0.20 and 5.10 卤0.35, respectively. The difference was statistically significant (P 0.05). The content of c GMP in the two groups was 0. 23 卤0. 02 卤0. 06 卤0.01)pg MGG ~ (-1), and the PKG content in these two groups was 0. 21 卤0. 02 卤0. 10 卤0.01)pg MGG ~ (-1), respectively. There were significant differences between the two groups (all P 0. 05). The expression of MHC in experimental group and model group was 1.55 卤0.161.1.15 卤0.22, respectively, and the expression of Tropnin1 in these two groups was 1.32 卤0.080.88 卤0.08, respectively. The expression of Desmin in these two groups was 1.33 卤0.51and 1.10 卤0.04, respectively. The difference was statistically significant (all P 0.05). Conclusion PDE5shRNA may reduce myocardial atrophy and myocardial fibrosis induced by adriamycin by activating c GMP/PKG pathway and improve left ventricular dysfunction.
【作者单位】：大连大学附属中山医院心内科;齐齐哈尔市第一医院心内科;
【基金】：齐齐哈尔市科学技术局计划基金资助项目
【分类号】：R965

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