<em>FrzA</em>基因转导干预缺血性心力衰竭小鼠心肌Wnt信号通路的研究

中国生物工程杂志

2013, Vol. 33

Issue (7): 13-17

研究报告

FrzA基因转导干预缺血性心力衰竭小鼠心肌Wnt信号通路的研究

沈鑫, 马依彤, 杨毅宁, 刘芬, 于子翔, 陈邦党, 陈铀

新疆医科大学第一附属医院乌鲁木齐 830000

Cardiac Transfection of AAV9-FrzA Gene Intervene Wnt Signal Pathway in Ischemic Heart Failure Mice

SHEN Xin, MA Yi-tong, YANG Yi-ning, LIU Fen, YU Zi-Xiang, CHEN Bang-dang, CHEN You

First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China

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摘要： 目的:研究重组9型腺相关病毒(recombinant adeno-associated virus serotype 9,rAAV9)携带FrzA基因转导干预缺血性心衰小鼠心肌Wnt信号通路的可行性,为基因治疗心力衰竭提供新的思路。方法:选择3月龄雄性C57BL/6J小鼠共130只,随机分为空白组(n=10),心衰组(n=40),心衰+空病毒(rAAV9-GFP)注射组(n=40),心衰+rAAV9-FrzA组(n=40),采用结扎左冠状动脉定量控制心梗面积,于术后2周行心脏超声评各组心功能变化,再经尾静脉注射已稀释好的病毒,28d后处死小鼠取心脏标本,RT-PCR检测心肌目的基因FrzA以及 Dvl-1,β-catenin的表达;Western blot检测心肌Wnt信号通路关键分子Dvl-1,GSK3β,p-GSK3β,β-catenin的表达。结果: 与空白组相比,成功建立心衰模型后小鼠心功能均不同程度降低(P<0.05);FrzA组与心衰组相比,心功能明显改善(P<0.05);经尾静脉注射可成功将rAAV9-FrzA导入小鼠体内,并且目的基因FrzA在心肌组织中高表达(P<0.05);心衰小鼠心肌中Wnt信号通路关键分子Dvl-1,p-GSK3β,β-catenin的表达显著升高(P<0.05),FrzA转导后小鼠心肌中Wnt信号通路中关键分子Dvl-1,p-GSK3β,β-catenin表达均降低(P<0.05)结论: 利用rAAV9-FrzA转导缺血性心衰小鼠可以有效的干预心肌Wnt信号通路,抑制其活性,为基因治疗缺血性心衰提供了新的思路。

关键词： 重组9型腺相关病毒; FrzA; 心力衰竭; Wnt信号通路

Abstract: Objective: To evaluate the feasibility of recombinant adeno-associated virus serotype 9 carrying FrzA Gene transfection to ischemic heart failure mice to intervene Wnt signal pathway. Methods: 130 of male C57BL/6J mice were divided into sham group(n=10), heart failure(HF) group(n=40), HF+rAAV9-GFP group(n=40), HF+rAAV9-FrzA group(n=40). Ligation of left coronary artery to create HF model and echocardiography was performed after 2 weeks. Then, mice were received tail-vein injection of either rAAV9-GFP or rAAV9-FrzA. After 28 days, the mice were used for detecting expression of FrzA, Dvl-1, GSK3β, p-GSK3β and β-catenin by RT-PCR or Western Blot. Result: rAAV9-FrzA was successfully transfected and expressed in mouse heart. Compared with sham group, heart failure could significantly increase the expression of Dvl-1, p-GSK3β and β-catenin(P<0.05); Compared with HF/HF+rAAV9-GFP groups, rAAV9-FrzA could significantly decrease the expression of Dvl-1, p-GSK3β and β-catenin(P<0.05).Conclusion: rAAV9-FrzA could effectively inhibit the Wnt signal pathway in ischemic heart failure mice, which may support a new idea for gene therpy of heart failure.

Key words: Recombinant adeno-associated virus serotype 9 FrzA Heart failure Wnt signal pathway

收稿日期: 2012-12-18 出版日期: 2013-07-25

ZTFLH:

Q342

基金资助: 国家自然科学基金(81060025);高等学校博士学科点专项科研基金(20106517110002);国家"973"计划前期研究专项(2010CB535013)资助项目

通讯作者: 马依彤 E-mail: myt-xj@163.com

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引用本文:

沈鑫, 马依彤, 杨毅宁, 刘芬, 于子翔, 陈邦党, 陈铀. FrzA基因转导干预缺血性心力衰竭小鼠心肌Wnt信号通路的研究[J]. 中国生物工程杂志, 2013, 33(7): 13-17.

SHEN Xin, MA Yi-tong, YANG Yi-ning, LIU Fen, YU Zi-Xiang, CHEN Bang-dang, CHEN You. Cardiac Transfection of AAV9-FrzA Gene Intervene Wnt Signal Pathway in Ischemic Heart Failure Mice. China Biotechnology, 2013, 33(7): 13-17.

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I7/13

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