一种不依赖呼吸机的小鼠心肌内注射技术的建立
投稿时间:2015-09-11  修订日期:2015-10-12  点此下载全文
引用本文:张逸杰,戴明彦,曹权,陈俊婷,包明威.一种不依赖呼吸机的小鼠心肌内注射技术的建立[J].医学研究杂志,2016,45(4):32-36
DOI: 10.11969/j.issn.1673-548X.2016.04.010
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作者单位E-mail
张逸杰 430060 武汉大学人民医院心内科、武汉大学心血管病研究所  
戴明彦 430060 武汉大学人民医院心内科、武汉大学心血管病研究所  
曹权 430060 武汉大学人民医院心内科、武汉大学心血管病研究所  
陈俊婷 430060 武汉大学人民医院心内科、武汉大学心血管病研究所  
包明威 430060 武汉大学人民医院心内科、武汉大学心血管病研究所 mbao@whu.edu.cn 
基金项目:国家自然科学基金资助项目(面上项目)(81570460);国家自然科学基金青年科学基金资助项目(81500668);中央高校基本科研业务费专项基金资助项目(2042014kf0172)
中文摘要:目的 建立一种简洁高效、不依赖辅助通气的小鼠心肌内注射的实验方法,并浅析操作细节及技巧。方法 44只6周龄的雄性C57BL/6J小鼠,随机分为实验组(22只)和对照组(22只)。麻醉后于左侧第4肋间隙挤出心脏,向左心室壁内注射靶向CPT-1b(实验组)或靶向无关基因(对照组)的重组慢病毒。10周后,取心脏及后肢骨骼肌,行RT-PCR及Western blot法,检测不同组织中CPT-1b的mRNA及蛋白表达水平。结果 至16周龄,小鼠存活率90.9%。实验组小鼠心肌中CPT-1b的表达显著低于对照组,而骨骼肌中CPT-1b的表达在两组间没有统计学差异。结论 心肌内注射技术所介导的RNA干扰高效、特异地下调了心肌组织中目标基因的表达,该技术是一种较好的将外源物质定向导入心肌组织的方法。
中文关键词:小鼠  心肌内注射  RNA干扰  动物模型  手术效率
 
Establishment of a Murine Model of Intramyocardial Injection Without a Ventilator
Abstract:Objective To establish a rapid and efficient murine model of intramyocardial injection without assisted ventilation, and comprehensively describe its details. Methods Six-week-old male C57BL/6J mice were randomized into experimental group (CPT group) and control group (Con group), and each group contained 22 animals. After deeply anesthetized, mice were subjected to small left thoracotomy followed by temporary cardio-exteriorization. Recombinant lentiviral vectors carrying short hairpin RNAs targeting murine carnitine palmitoyltransferase-1b (CPT-1b, for CPT group) or irrelevant gene (for Con group) were injected into left ventricular wall directly. Ten weeks later, myocardium and skeletal muscle were obtained. CPT-1b mRNA and protein levels expressed in different tissues were measured by RT-PCR and Western blot. Results The survival rate of all animals was 90.9% till 16 weeks of age. CPT-1b expression in the myocardium was significantly lowered in CPT group compared to Con group, while its expression in skeletal muscle was indistinguishable between the two groups. Conclusion Intramyocardial injection-mediated RNA interference effectively and specifically down-regulated the expression of targeted gene in the myocardium. This new technology represents a preferable approach to introduce exogenous substance into the myocardium of a mouse.
keywords:Mouse  Intramyocardial injection  RNA interference  Animal models  Surgical efficiency
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