张彦燕,黄勇攀,石刚刚
ZHANG Yanyan,HUANG Yongpan,SHI Ganggang

• 1:贵阳医学院药理学教研室
• 2:汕头大学医学院药理学教研室

摘要(Abstract)：

目的:建立适应于膜片钳实验的大鼠单个心室肌细胞的分离方法,探讨L-型钙通道在大鼠心肌电活动中的作用。方法:采用改进Langendorff装置,酶解法分离单个心室肌细胞,应用全细胞膜片钳技术记录心室肌细胞L-型钙电流及维拉帕米的作用。结果:该法分离到的心肌细胞存活率达80%~90%,高倍镜下细胞呈杆状,边缘清楚,表面光滑,纹理清晰;成功记录到典型L-型钙电流(ICa-L),给予维拉帕米(0.3、3、30μmol/L)后呈浓度依赖性抑制钙电流,其抑制率分别为(44.35±5.52)%,(61.06±2.99)%,(71.88±4.45)%。结论:改进心肌细胞的分离方法可获得大量耐钙心室肌细胞,这些细胞具有正常的电生理特性。
Objective: To establish a method for isolating single cardiac myocyte for patch clamp experiment,and to explore the role of L-type calcium channels in cardiac electrical activity in rats.Methods: The ventricular myocytes were isolated by using enzymolysis method with improved Langendorff apparatus. L-type calcium current and the effect of verapmil were recorded by using whole-cell patch clamp technique. Results: The survival rate of myocardial cells was 80% ~ 90%. The cells were rod-shaped with clear edge,smooth surface,clear texture at high magnification. L-type calcium current was recorded successfully. Verapamil inhibited the L-type calcium current in a concentrationdependent manner. The inhibitory rate of 0. 3,3,30 μmol /L verapamil were( 44. 35 ± 5. 52) %,( 61.06 ±2. 99) %,( 71. 88 ±4. 45) % respectively. Conclusions: The improved isolating method can obtain a large number of single cardiac myocytes with strong calcium resistance ability and normal electrophysiological properties.

关键词(KeyWords)： 心肌;细胞分离;膜片钳术;L-型钙电流;膜电位
myocardium;cell separation;patch clamp technique;L-type calcium current;membrane potential

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81102664);; 贵州省科学技术基金[黔科合J字(2011)2285号,(2013)2050号];; 贵州省卫生厅科学技术基金(gzwkj2011-1-019)

作者(Author): 张彦燕,黄勇攀,石刚刚
ZHANG Yanyan,HUANG Yongpan,SHI Ganggang

DOI: 10.19367/j.cnki.1000-2707.2014.05.007

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