单个人心房肌细胞的分离、鉴定及影响因素分析  

Isloation and identification of single human atrial myocytes and its effect fators

作　　者：刘金平[1] 孙宗全[1] 张昌伟[1] 刘坤[1] 高翔[1] 水志刚[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院,湖北武汉430022

出　　处：《山东医药》2008年第29期7-9,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30570729);湖北省卫生厅科研基金资助项目(JX3B08)

摘　　要：目的探讨分离、鉴定单个人心房肌细胞的最佳方法及其影响因素。方法人心房肌组织取自5例先天性心脏病（PCHD）、8例冠心病（CHD）、15例瓣膜性心脏病（VHD）手术患者，采用两步酶解法分离制备心房肌细胞，光学显微镜下观察细胞形态，全细胞膜片钳技术记录乙酰胆碱敏感钾电流（Ik,Ach）情况。结果两步酶解法分离所得杆状、横纹清楚、折光性好、静止、贴壁良好的心肌细胞产率为30％～50％。PCHD记录Ik,Ach成功率最高（9／10个细胞），其次为CHD（6／10个细胞），VHD最低（3／10个细胞）。结论两步酶解法可用于人心房肌细胞的分离，全细胞膜片钳技术有助于细胞活性的鉴定，其分离数量和活性受酶解情况、年龄、病种等因素影响。Objective To explore the optium method of isolating single human atrial myocytes and its effect factors. Methods Single human atrial myocytes from patients with various ages and heart diseases -pediatric congenital heart diseases （PCHD） , coronary heart diseases （CHD） , valvular heart diseases（VHD） were prepared with a two-step enzymatic method. These myocytes were observed by light microscope to show the shape, striation, refraction, quiescence and adherence and tested by the whole cell patch clamp technique to record muscarinic-gated atrial potassium currents （IK, Ach）. Results The viability of the rod-shaped, striated, refractive,quiescent and adherent myocytes ranged from 30% to 50%. IK ,Ach currents were recorded the highest in the myocytes from patients with PCHD （9/10 cells）, second from patients with CHD （6/10 cells） , third from pa＇ients with VHD（3/10 cells）. Conclusion Two-step enzymatic method can be used to the isolation of single human atnal myocytes, the whole cell patch clamp technique is helpful to the identification of cell activity. The viability of single human atrial myocytes can be affected by enzymes disassemble, ages and kind of diseases.

关 键 词：心房肌细胞 膜片钳 两步酶解法 影响因素 

分 类 号：R318[医药卫生—生物医学工程]