【作者】 商立军；

【导师】 臧益民； 董秀珍； 王四旺； 周士胜； 谢安；

【作者基本信息】 第四军医大学 ， 生理学， 2000， 博士

【摘要】 CGRP是一种重要的心血管系统调节肽，与许多心血管疾病密切相关。近年来研究发现，CGRP具有明显的抗心律失常作用，但国内外文献报道较少，且其抗心律失常作用的电生理机制和离子机制尚不清楚。缺血性和缺血再灌注性心律失常在临床上十分常见且后果严重，但目前治疗效果欠佳，故寻求一种有效的治疗方法是目前研究的热点。本课题运用膜片钳和激光扫描共聚焦技术来探讨CGRP对正常及模拟缺血、缺氧心肌细胞钙通道的作用及其电生理机制，为其广泛应用于临床提供理论和实验依据。同时，本课题利用计算机重建技术，在心室肌细胞模型及自身实验的基础上，对钠、钾、钙通道动力学过程及电流进行重建，以弥补动物实验的不足，尝试通过定量化的研究，为解释动物实验结果提供理论依据的有效途径。 主要研究内容包括： 1．掌握心肌细胞培养技术，研究CGRP对培养心肌细胞的直接作用。 2．掌握激光扫描共聚焦技术，讨论CGRP对急性分离心肌细胞在模拟缺血、缺氧状态下胞内钙的影响。 3．掌握急性分离心室肌细胞全细胞膜片钳实验技术，掌握计算机记录、分析系统的使用。 4．在电压钳状态下记录心肌细胞L型钙电流，讨论CGRP对正常及模拟缺血缺氧心肌钙电流的作用。 5．建立能模拟心肌细胞钠、钾、钙通道的动力学过程及电流的数学模型，并进行仿真研究。 通过以上研究，得到如下主要结果和结论： 1．1×10^-9mol·L^-1和1×10^-8mol·L^-1浓度的CGRP对心肌细胞无毒性作用，且能使心肌细胞的搏动频率加快，呈现正性变时性和正性变力性效更多还原

【Abstract】 Calcium Gene-Related Peptide (CGRP) is an important cardiovascular regulating peptide and has close connection with many cardiovascular diseases. In recent studies, CGRP showed its notable protective effects on arrhythmias. But there is still few reports on the effect. Especially the electrophysiological and ionic machenisms of this effect are unclear. Ischemia and reperfusion arrhythmias are very common with serious consequences on clinical manifestation and therapies for them are not satisfied. Therefore there comes urgency for finding an effective method to cure them. By using the patch clamp technique and laser scanning confocal technique, this study discussed the effects of CGRP on cardiac calcium channel on normal condition as well as on imitating ischemia and hypoxia conditions. And this study tried to discuss the electrophysiolodical ionic mechanisms of these effects so as to provide some theoretical and experimental proofs for wider clinical application. Meanwhile this study tried to reconstruct sodium, potassium and calcium channel dynamic courses and currents based on ventricular myocytes mathematical models and my personal experiments by using computer construction technique. The study could remedy the defect of animal experiments and open an effective way for providing theoretical proof for explaining the data from animal experiments by quantity study.The experimental designs were as follows:1. To grasp the culture technique of cardiomyocytes and study the direct更多还原