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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(P<0.05),梗死周边心肌细胞凋亡明显减少(P<0.05),实验组小鼠心肌PDE5表达明显减少,c GMP和PKG表达明显上调(P<0.05)。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。
Objective To investigate the effect of PDE5 shRNA targeting short PDE5 shRNA on early apoptosis of acute myocardial infarction in mice. Methods The model of myocardial infarction in mice was established by ligation of the left anterior descending coronary artery in mice and randomly divided into experimental group (n = 10). The recombinant adenovirus vector carrying PDE5 shRNA was injected into myocardial infarction area and peripheral area and the control group. The common adenovirus vector inserted into PDE5 shRNA was injected into the myocardial infarction area and peripheral area. One week later, immunohistochemistry and TUNEL were used to detect the apoptosis of infarcted and infarcted cells. The levels of cGMP and protein kinase G (cGMP) were detected by enzyme-linked immunosorbent assay (ELISA) PKG) expression, Western blot analysis to detect the expression of PDE5 in each group. Results Compared with the control group, the apoptosis of infarcted area and surrounding area of infarcted area in experimental group was significantly decreased (P <0.05), and the apoptosis of myocardial cells in infarcted area was significantly decreased (P <0.05) Myocardial PDE5 expression was significantly reduced, cGMP and PKG expression was significantly increased (P <0.05). Conclusion PDE5 shRNA can significantly reduce the apoptosis in myocardial infarction and surrounding infarcts and myocardial cells in non-infarcted area, which may be related to the up-regulation of c-GMP and PKG expression.