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目的:探讨磷酸二酯酶5(PDE5)shRNA对小鼠急性心梗后早期细胞凋亡的影响。方法:通过结扎小鼠左冠状动脉前降支建立小鼠心肌梗死模型,并将建立好的模型随机分为实验组(n=10):将携带有抑制PDE5的shRNA(PDE5 shRNA)重组腺病毒载体对小鼠心肌组织多部位注射;模型组(n=10):将没有插入PDE5 shRNA的普通腺病毒载体对小鼠心肌组织多部位注射。1周后,进行免疫组化和TUNEL分析检测梗死及梗死周边细胞凋亡情况,ELISA法检测环磷酸鸟苷(cyclic guanosine monophosphate,c GMP)和蛋白激酶G(protein kinase G,PKG)的表达,免疫蛋白印迹分析检测各组磷酸二酯酶5(phosphodiesterase5,PDE5)的表达情况。结果:心梗1周后,和模型组相比较,实验组小鼠梗死区及梗死周边区域细胞凋亡明显减少(P<0.05),实验组小鼠心肌PDE5表达明显减少,cGMP和PKG表达明显上调(P<0.05)。结论:PDE5 shRNA可以明显减少梗死区及梗死周边区细胞凋亡,可能与上调cGMP和PKG的表达密切相关。
Objective: To investigate the effect of phosphodiesterase 5 (PDE5) shRNA on early apoptotic cells in acute myocardial infarction mice. Methods: The model of myocardial infarction in mice was established by ligation of left anterior descending coronary artery in mice. The established model was randomly divided into experimental group (n = 10): recombinant adenovirus carrying PDE5 shRNA (PDE5 shRNA) The vector was injected into the myocardium of mice at multiple sites. The model group (n = 10): The common adenovirus vector without PDE5 shRNA was injected into the myocardium of mice. One week later, immunohistochemistry and TUNEL assay were used to detect the apoptosis of infarcted and infarcted cells. The expression of cyclic guanosine monophosphate (cGMP) and protein kinase G (PKG) The expression of phosphodiesterase 5 (PDE5) in each group was detected by Western blot analysis. Results: After 1 week myocardial infarction, compared with the model group, the apoptotic rate in the experimental group and the peripheral infarction group were significantly decreased (P <0.05), and the expression of cGMP and PKG in the experimental group was significantly decreased (P <0.05). Conclusion: PDE5 shRNA can significantly reduce apoptosis in the infarct area and peripheral infarction area, which may be closely related to the up-regulation of cGMP and PKG expression.