【摘要】 目的 探究腺相关病毒不同转染策略构建大鼠心脏光遗传学模型的可行性与有效性。方法 20只SD大鼠，随机分为4组：心肌注射组、心包注射组、心外膜滴涂组、正常对照组，每组5只，分别心肌注射、心包注射、心外膜滴涂rAAV9-CAG-hChR2(H134R)-eYFP,正常对照组只开胸。造模4周后，超声测量4组大鼠左室射血分数及左室短轴缩短率；记录体表心电图，测量RR间期、PR间期和QRS时限。开胸进行473 nm波长蓝光照明实验，根据光输出信号与心电图判断心脏节律是否被光脉冲夺获。对心脏组织石蜡切片进行HE染色和荧光观察。结果 心肌注射组、心包注射组、心外膜滴涂组大鼠与正常对照组大鼠相比，心电参数无显著差异(P>0.05),左室射血分数及左室短轴缩短率无显著差异(P>0.05)。用一定频率和功率的473 nm蓝光照射心肌注射部位或心脏，能夺获心肌注射组和心包注射组心脏的节律，但不能夺获心外膜滴涂组。HE染色显示心肌注射组注射部位心肌纤维排列紊乱、炎症细胞浸润。荧光显示心肌注射组、心包注射组、心包滴涂组心室切片均有ChR2(H134R)转染。结论 心包注射途径能有效构建大鼠心脏光遗传学模型。更多还原

【Abstract】 Objective To investigate the feasibility and effectiveness of different transfection strategies of adeno-associated virus to construct a rat cardiac optogenetic model.Methods A total of 20 SD rats were purchased and randomly divided into four groups: myocardial injection group, pericardial injection group, epicardial drip coating group and normal control group.The rats(n=5) were injected with rAAV9-CAG-hChR2(H134R)-eYFP by myocardial injection, pericardial injection and epicardial drip coating respectively.After Four weeks, left ventricular ejection fractions(LVEF) and left ventricular fractional shortening(LVFS) of all rats were measured by ultrasound; record the body surface ECG to measure the RR interval, PR interval and QRS wave time frame.Open-chest light experiments with 473 nm blue light were performed to determine whether the heart rhythm was captured by the light pulse based on the light output signal and the body surface ECG.Results The rats in the myocardial injection group, pericardial injection group and epicardial drip coating group, compared with the rats in the normal control group, showed no significant differences in ECG parameters(P>0.05) and no significant differences in LVEF and LVFS(P>0.05).The rhythm of myocardial injection and pericardial injection groups could be captured by irradiating the myocardial injection site or heart with 473 nm blue light of certain frequency and power, but not the epicardial drip coating group.HE staining showed disorganized myocardial fiber arrangement and inflammatory cell infiltration at the injection site in the myocardial injection group.Fluorescence showed ChR2(H134R) transfection in ventricular sections of the myocardial injection group, pericardial injection group, and pericardial drip coating group.Conclusion The pericardial injection can effectively construct cardiac optogenetics model.更多还原