利用光遗传学技术实施终止室性心律失常的实验研究  

Investigation on the efficacy of ventricular arrhythmia defibrillation by optogenetic technique

作　　者：程玥 李剑怡 饶盼盼[1] 刘华芬[1] 王晞[1] CHENG Yue;LI Jian-yi;RAO Pan-pan;LIU Hua-fen;WANG Xi(Department of Cardiology,Renmin Hospital of Wuhan University,Cardiovascular Research Institute of Wuhan University,Hubei Key Laboratory of Cardiology,Wuhan 430060,Hubei,China)

机构地区：[1]武汉大学人民医院心内科武汉大学心血管病研究所心血管病湖北省重点实验室,湖北武汉430060

出　　处：《中国心脏起搏与心电生理杂志》2020年第4期376-381,共6页Chinese Journal of Cardiac Pacing and Electrophysiology

摘　　要：目的探讨基于光遗传学技术的程控光照终止大鼠室性心律失常的有效性及其影响因素。方法将SD大鼠18只随机等分成3组:ChR2光照组,ChR2未光照组,空载病毒组,前两组动物经颈静脉注射100μl光敏蛋白载体病毒pAAV-CAG-hChR2(H134R)-mCherry,空载病毒组注射100μl空载病毒pAAV-CAG-mCherry。8周后,所有大鼠均麻醉后开胸充分暴露心脏,结扎冠状动脉左前降支后给予Burst电刺激(10 V,50 Hz,100个脉冲,持续2 s)诱发心室颤动或室性心动过速,ChR2光照组和空载病毒组分别给予4个间隔1 s的脉冲宽度为1 s的473 nm蓝光照射心外膜表面,ChR2未光照组不进行光照,观察光照终止的有效性,同时改变光功率密度(2、4、8、10、20倍阈值)观察不同光功率密度对终止效率的影响。对心脏组织石蜡切片进行HE染色和荧光观察。结果 ChR2光照组的终止成功率为98.0%±2.0%,明显高于ChR2未光照组(51.7%±6.0%)和空载病毒光照组(57.2%±6.8%)。功率密度依次增加(2、4、8、10以及20倍功率密度阈值),终止率也逐渐增加,分别为64.0%±6.8%、70.0%±7.1%、74.0%±8.7%、88.0%±3.7%和98.0%±2.0%。HE染色显示急性心肌梗死存在,石蜡切片红色荧光显示ChR2光照组、ChR2未光照组转染目标光敏感蛋白ChR2成功。而空载病毒组未表达ChR2。结论表达了ChR2的心脏发生室性心律失常,可被光遗传学技术有效终止,且终止成功率与光照功率密度相关。Objective To demonstrate the effectiveness of optogenetics in the termination of ventricular arrhythmia in adult rats with acute myocardial infarction and the effect of optical intensity density on defibrillation efficiency. Methods The experimental animals were randomly divided into 3 groups: 6 in the ChR2 illumination group, 6 in the ChR2 unilluminated group, and 6 in the virus without anti-Smad group. The first and second groups of SD rats were injected with 100 μl of pAAV-CAG-hChR2(H134 R)-mCherry via the jugular vein. The third group was injected with 100 μl of empty virus pAAV-CAG-mCherry. After 8 weeks, the rats were completely anesthetized and then opened the chest, the heart was fully exposed. The left anterior descending coronary artery was ligated to establish an acute myocardial infarction, and the light output signal and the surface electrocardiogram were simultaneously recorded. Ventricular arrhythmia was induced by Burst stimulation(10 V, 50 Hz, 100 pulses, lasting 2 s), and four pulses with width of 1 s at 1 s interval was irradiated onto the epicardial surface for optogenetic defibrillation to evaluate the effectiveness of defibrillation. At the same time, the optical intensity density(2,4,8,10,20 times of the threshold intensity density) was changed to observe the effect of different optical intensity densities on the defibrillation efficiency. HE staining and fluorescence section observation were performed on paraffin sections of heart tissue Results The defibrillation rates of the ChR2 illumination group, the ChR2 unilluminated group and the virus without anti-Smad group were 98.0%±2.0%, 51.7%±6.0% and 57.2%±6.8 %, respectively, confirmed the effectiveness of optogenetics in the application of cardiac defibrillation. The intensity density increased sequentially (2,4,8,10 and 20 xintensity density threshold),and the defibrillation rate also increased gradually,which is64.0%±6.8%,70.0%±7.1%,74.0%±8.7%,88.0%±3.7% and 98.0% ±2.0%.HE staining showed the presence of acute myocardial infarc

关 键 词：光遗传学 心室颤动 室性心动过速 光敏蛋白 腺相关病毒 

分 类 号：R541.7[医药卫生—心血管疾病]