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目的 探讨重组9型腺相关病毒(rAAV-9)介导R65核酶基因转导对小鼠急性心肌梗塞(AMI)后Wnt信号通路的影响。方法将3个月龄雄性C57/BL小鼠60只随机分为2组:AMI+R65组每日尾静脉注射携带增强型绿色荧光蛋白(enhanced green fluorescent protein,eGFP)报告基因及R65核酶基因的rAAV9转染小鼠。AMI组每日按同等剂量注射rAAV9-eGFP,21d后结扎左冠状动脉,建立小鼠AMI模型。所有小鼠按心梗时间分3、7、14 d组,比较R65核酶干预后p65、糖原合成激酶(GSK)-3β总水平及磷酸化水平变化,β-联接蛋白(catenin)及缝隙通道连接蛋白(connexin)43表达。结果与AMI组相比,R65组p65表达降低,GSK-3β磷酸化比值3 d时降低,7 d及14 d时升高(P值均为0.000);AMI+R65组-βcatenin表达3 d时低于AMI组,7 d时高于AMI组(P值均为0.000);AMI+R65组connexin43表达明显低于AMI组,差异有统计学意义(P=0.000)。结论通过R65核酶过表达抑制NF-κB活性后Wnt信号通路相关蛋白活性发生变化,说明急性心肌梗塞后NF-κB信号通路与Wnt信号通路之间也存在“crosstalk”。
Objective To investigate the effect of recombinant adenovirus-9 (rAAV-9) on the Wnt signaling pathway after acute myocardial infarction (AMI) in mice mediated by R65 ribozyme gene transduction. Methods 60 male C57 / BL 3 months old mice were randomly divided into 2 groups: AMI + R65 group was injected daily with tail vein injection of enhanced green fluorescent protein (eGFP) reporter gene and R65 ribozyme gene Of rAAV9-transfected mice. The rats in AMI group were injected rAAV9-eGFP at the same dose daily, and the left coronary artery was ligated after 21 days. The AMI model of mice was established. All the mice were divided into 3, 7 and 14 d groups according to myocardial infarction time. The levels of p65, GSK-3β, phosphorylation level, β-catenin and gap Channel connexin 43 expression. Results Compared with AMI group, the expression of p65 was decreased in R65 group and the phosphorylation of GSK-3β was decreased at 3 d and increased at 7 d and 14 d (P <0.0001 respectively); when the expression of β-catenin in AMI + R65 group was 3 d The level of connexin43 in AMI + R65 group was significantly lower than that in AMI group (P = 0.000). Conclusions The activity of Wnt signaling pathway related protein changes after NF-κB activation is inhibited by R65 ribozyme, indicating that there exists “crosstalk” between NF-κB and Wnt signaling pathway after acute myocardial infarction.