靶向磷酸二酯酶5的短发夹RNA重组腺病毒载体对小鼠急性心梗后早期细胞凋亡的影响

发布时间：2019-02-09 14:39

【摘要】：目的探讨靶向持续抑制磷酸二酯酶5(PDE5)的短发夹RNA(shRNA)[PDE5 shRNA]重组腺病毒载体对小鼠急性心梗后早期细胞凋亡的影响。方法结扎小鼠左冠状动脉前降支建立小鼠心肌梗死模型,随机分为实验组10例,将携带有PDE5 shRNA重组腺病毒载体注射到心肌梗死区及周边区和对照组10例,将没有插入PDE5 shRNA的普通腺病毒载体注射到心肌梗死区及周边区。1周后,进行免疫组化和原位末端标记分析(TUNEL)检测梗死及梗死周边细胞凋亡情况,酶联免疫吸附实验(ELISA法)检测环磷酸鸟苷(c GMP)和蛋白激酶G(PKG)的表达,蛋白印迹分析检测各组PDE5的表达情况。结果心梗1周后,和对照组相比较,实验组小鼠梗死区及梗死周围区域细胞凋亡明显减少(P0.05),梗死周边心肌细胞凋亡明显减少(P0.05),实验组小鼠心肌PDE5表达明显减少,c GMP和PKG表达明显上调(P0.05)。结论 PDE5 shRNA的干预可以明显减少心肌梗死及梗死周边区细胞凋亡和非梗死区心肌细胞凋亡,可能与上调c GMP和PKG的表达密切相关。
[Abstract]:Objective to investigate the effect of short hairpin RNA (shRNA) [PDE5 shRNA] recombinant adenovirus vector targeting phosphodiesterase 5 (PDE5) on early apoptosis after acute myocardial infarction in mice. Methods the myocardial infarction model was established by ligating the anterior descending branch of left coronary artery in mice. The model was randomly divided into experimental group (n = 10) and control group (n = 10). The recombinant adenovirus carrying PDE5 shRNA was injected into myocardial infarction area and peripheral area and control group (n = 10). The normal adenovirus vector not inserted into PDE5 shRNA was injected into the myocardial infarction area and peripheral area. 1 week later, the apoptosis of infarct and periinfarction cells was detected by immunohistochemistry and in situ end labeling analysis (TUNEL). Enzyme linked immunosorbent assay (ELISA) was used to detect the expression of cyclic guanosine monophosphate (c GMP) and protein kinase G (PKG) (PKK). Western blot analysis was used to detect the expression of PDE5 in each group. Results after one week of myocardial infarction, compared with the control group, the apoptosis of myocardial cells in the experimental group was significantly decreased (P0.05), and that in the infarcted area was significantly decreased (P0.05). The expression of PDE5 in myocardium of experimental group was significantly decreased, the expression of, c GMP and PKG was significantly up-regulated (P0.05). Conclusion the intervention of PDE5 shRNA can significantly reduce the apoptosis of myocardial infarction and periinfarction area and myocardial cell apoptosis in non-infarct area, which may be related to the up-regulation of c GMP and PKG expression.
【作者单位】：齐齐哈尔市第一医院干部病房;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;齐齐哈尔市第一医院科教部;
【基金】：国家自然科学基金资助项目(81170187) 教育部留学回国人员科研启动基金资助项目[教外司留(2013)693号]
【分类号】：R542.22

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