SHR大鼠心室肌细胞分离方法改进及其L-型钙电流的观察

Improved method for isolation of SHR rats ventricular myocytes and observation of L-type calcium current

文章探索建立简单可靠的SHR大鼠心室肌细胞的分离方法,提高细胞存活率和膜片钳试验成功率。采用Langendorff灌流装置,离体心脏逆向灌流,分离单个SHR大鼠心室肌细胞。使用膜片钳全细胞记录技术,记录大鼠心室肌细胞L-型钙电流。通过对细胞的观察发现,细胞呈长杆状,横纹清晰,细胞膜结构完整。膜片钳全细胞记录到典型的L-型钙电流,激活电位-40 mV,峰值电位0 mV,反转电位+50～+60 mV,呈现出电压依赖性。观察到的L-型钙电流可被钙通道阻滞剂维拉帕米抑制。结果表明,采用的大鼠心肌细胞分离方法可分离制备出适合膜片钳电生理观察的心肌细胞标本。

A simple and reliable SHR rat ventricular myocytes isolation method was explored and established to improve cell survival and the success rate of patch-clamp experiments. In present study, the isolation method of rats ventricular myocytes was conducted to investigate for the patch clamp experiment and observed the L-type calcium channel current. The Langendorff perfusion method was used to isolate the ventricular myocytes from SHR rats. The whole-cell patch clamp technique was used to record the calcium current （Ic^L） of beingisolated ventricular myocytes from SHR rats. The observation of cells showed that this method was well-constructed, rod-like and the cross striation could be observed clearly. The typical L-type calcium current was recorded by the whole-cell patch clamp recording. The activation potential was -40 mV; peak potential was 0 mV; reversal potential was ＋50-＋60 mV; Meamwhile it was in the voltage-dependent manner. The L-type calcium current observed could be inhibited by the calcium channel blocker-Verapamil. The rats ventricular myocyte samples which were isolated and prepared by this method would be suitable for the patch clamp electrophysiological observation.