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SERCA2a 基因转导过表达增强犬心肌细胞收缩功能

陈礼斌, 宫海滨, 刘颖, 王振全, 吕茜

东南大学医学院临床医学系，南京210009；徐州市心血管病研究所，徐州市中心医院，徐州 221009

摘要

本研究旨在探讨心肌肌浆网Ca2+-ATP 酶2a (sarcoplasmic reticulum Ca2+-ATPase 2a, SERCA2a)基因过表达对正常犬离体心肌细胞收缩功能的影响。通过胶原酶消化法分离犬心肌细胞，将分离的心肌细胞分为未转染组、空载体组、SERCA2a转染组，转染载体为含绿色荧光蛋白基因的重组腺病毒载体，转染后均培养48 h 。利用免疫印迹法检测各组心肌细胞SERCA2a 蛋白表达水平，并通过单细胞收缩动态边缘检测系统测定各组心肌细胞收缩功能的改变。结果显示，与未转染组相比，SERCA2a 转染组SERCA2a 蛋白表达水平明显升高，基础状态心肌收缩百分比和药物刺激条件下心肌最大收缩百分比均明显升高，达到峰值收缩时间(TTP)和50% 舒张时间(R50)亦明显延长，差异具有显著性；而空载体组各项指标均无显著变化。以上结果提示，SERCA2a 基因转导过表达能够增强正常犬心肌细胞的收缩功能。

关键词： 肌浆网C a 2 +-ATP 酶; 犬; 腺病毒; 基因

分类号：R318.11

[Overexpression of SERCA2a by gene transfer enhances myocardial systolic function in canines.] [Ariticle in Chinese]

[Overexpression of SERCA2a by gene transfer enhances myocardial systolic function in canines.] [Ariticle in Chinese]

Department of Clinical Medicine, Southeast University, Nanjing 210009, China

Abstract

The present study is aimed to study the effect of sarcoplasmic reticulum Ca2+-ATPase 2a (SERCA2a) gene transfer on the contractile function of isolated cardiomyocytes of canines. The cardiomyocytes were isolated with collagenases. The isolated cardiac cells were divided into untransfected group, empty vector group and SERCA2a-transfected group. Recombinant adenovirus vector carrying enhanced green fluorescent protein gene was used for SERCA2a gene delivery. The expression of SERCA2a protein in cardiomyocytes was determined by Western blot. Contractile function of cardiomyocytes was measured with motion edge-detection system of single cell at 48 h after transfection. The results showed, compared with untransfected group, SERCA2a protein level, percentage of peak contraction amplitude under normal condition, percentages of peak contraction amplitude under Ca2+ or isoproterenol stimulation, time-to-peak contraction (TTP) and time-to-50% relaxation (R50) in SERCA2a-transfected group all increased significantly. While all the above indices in empty vector group did not show any differences with those in untransfected group. These results suggest that the overexpression of SERCA2a by gene transfer may enhance the contraction function of canine myocardial cells.

Key words: sarcoplasmic reticulum Ca2+-ATPase; canine; adenovirus; gene

收稿日期：2010-09-06　　录用日期：2010-11-11

通讯作者：宫海滨　　E-mail: ghbxzh@yahoo.com.cn

引用本文：

陈礼斌, 宫海滨, 刘颖, 王振全, 吕茜. SERCA2a 基因转导过表达增强犬心肌细胞收缩功能[J]. 生理学报 2010; 62 (6): 511-516.

[Overexpression of SERCA2a by gene transfer enhances myocardial systolic function in canines.] [Ariticle in Chinese] . [Overexpression of SERCA2a by gene transfer enhances myocardial systolic function in canines.] [Ariticle in Chinese] . Acta Physiol Sin 2010; 62 (6): 511-516 (in Chinese with English abstract).