重组9型腺相关病毒介导R65核酶基因转导对小鼠急性心肌梗塞后Wnt信号通路的影响  

Effects from transfection of recombinant adeno-associated virus serotype 9 with R65 ribozyme to Wnt signal pathway in mice post-infarction

作　　者：黄莺[1] 安尼瓦尔.阿不里孜 马依彤[1] 杨毅宁[1] 刘芬[1] 陈邦党[1] 李晓梅[1] 

机构地区：[1]新疆医科大学第一附属医院冠心病科,新疆乌鲁木齐830011

出　　处：《新疆医科大学学报》2011年第6期568-572,共5页Journal of Xinjiang Medical University

基　　金：国家重点基础研究发展(973计划)项目(2010CB535013);新疆医科大学第一附属医院科研奖励基金(2010YFY25)

摘　　要：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 evaluate the effects from the transfection of recombinant adeno-associated virus serotype 9 carrying enhanced green fluorescent protein（rAAV9-eGFP） and R65 ribozyme to Wnt signal pathway in mice post-infarction.Methods Choose 60 3-months C57/BL mice and divide them into 2 groups： AMI group were transfected with rAAV9-eGFP and AMI＋R65 group with rAAV9-eGFP-R65 by tail injection for 21 d then we set up acute myocardial infarction animal model by ligation of left coronary artery.They were divided into 3 subgroups according to time post-infarction： 3,7,14 d post infarction and detected expression of p65 by immunohistochemistry;total（t）and phosphorylation（p） level of glucogen synthesis kinase（GSK）-3β,β-catenin,connexin 43 by western-blot in left ventricular（LV）.Results Expression of p65 decreased in AMI＋R65 group（P0.05）.Compared with AMI group ratio of p/t-GSK-3β is higher in AMI＋R65 group in 3 d and lower in 7 d post infarction than it in AMI group（P value all 0.000）.Expression of β-catenin is lower in AMI＋R65 group in 3 d postinfarction,while it increased in AMI＋R65 group on 7 d postinfarction（P=0.000）.Expression of connexin 43 was much lower in AMI＋R65 group compared with AMI group（P=0.000.Conclusions Expression of Wnt signal pathway altered after we used R65 ribozyme to inhibit activities of NF-κB.That means ＂crosstalk＂ between Wnt and NF-κB existed post infarction might relate to p65 ability.

关 键 词：心肌梗死 NF-ΚB信号通路 WNT信号通路 

分 类 号：R34[医药卫生—基础医学] R-332