大鼠心室肌细胞的分离及其钙电流的观察  

Isolation Technique and Observation on Calcium Current of Ventricular Myocytes from Rat

作　　者：黄展勤[1] 高分飞[1] 石刚刚[1] 

机构地区：[1]汕头大学医学院药理教研室,广东汕头515041

出　　处：《汕头大学医学院学报》2003年第2期102-104,共3页Journal of Shantou University Medical College

基　　金：国家自然科学基金 ( 3 0 0 70 3 0 4);广东省科技计划项目 (C3 0 10 4);广东省自然科学基金重点项目 ( 62 12 3 5 )

摘　　要：目的 :探讨大鼠心肌细胞酶急性分离的方法 ,并观察大鼠心室肌细胞膜钙电流。方法 :采用Langendorff灌流装置 ,用含胶原酶P(0 12g L)的台式液 ,经主动脉逆行灌流心脏 ,分离心肌细胞 ;采用膜片钳全细胞记录的方法 ,观察记录大鼠心室肌细胞膜L_型钙电流。结果 :心肌细胞的存活率约 70 %～80 % ,形态呈长杆状 ,横纹清晰 ,膜良好。膜片钳全细胞记录出电压依赖性内向电流 ,其激活电位 - 30mV ,最大激活电位 0mV ,反转电位 5 0mV ,并且此内向电流可被钙通道阻滞剂维拉帕米抑制。结论 :用此法可分离出高存活率的心肌细胞 。Objective: To investigate the acute method to isolate s ingle ventricular myocyte from rat by the use of enzymatic dissociation and obse rve the calcium current in rat ventricular myocytes.Methods: Heart was perfused by Tyrode solution with collagenase P(0 12g/L) on the Langendorff appar atus. And the calcium current was observed by the use of patch clamp technique w hole cell recording. Results: With the enzymatic dissociation, the well_construc ted and rod_like cardiac myocytes were got. Under the depolarizing potential as -30 mV, the inward current was elicited. The peak activated potential was 0 mV. And the reversal potential was 50 mV. The inward current was inhibited by Verapa mil, a typical calcium channel antagonist. Conclusion: The procedure of isolatio n used in this study can produce large numbers of single cardiac myocytes with h igh survival rate and normal electrophysiological properties.

关 键 词：大鼠 心室肌细胞 分离 钙电流 观察 膜片钳 胶原酶 

分 类 号：R319[医药卫生—基础医学]