磷酸化组蛋白H3--检测心肌细胞有丝分裂指数的可靠指标  

PHOSPH-HISTONE H3——AN APPROACH TO DETECT THE MITOTIC INDEX OF CARDIOMYOCYTES

作　　者：邸菁[1] 张凌志[2] 柏树令[1] 

机构地区：[1]中国医科大学基础医学院人体解剖学教研室,沈阳110001 [2]中国医科大学附属盛京医院急诊科,沈阳110004

出　　处：《解剖学报》2008年第2期272-274,共3页Acta Anatomica Sinica

基　　金：辽宁省教育厅资助项目(051525)

摘　　要：目的采用磷酸化组蛋白H3鉴定心肌细胞有丝分裂,为研究心肌细胞增殖机制奠定形态学基础。方法对H9c2(2-1)大鼠心肌细胞进行培养,利用免疫荧光双重染色技术,用横纹肌肌动蛋白IgM单克隆抗体标记心肌细胞,用磷酸化组蛋白H3 IgG单克隆抗体标记有丝分裂的染色体,同时用Hochest 33342显示细胞核,荧光显微镜观察并摄片。结果心肌细胞胞质呈红色荧光,有丝分裂的染色体呈现不同形状的绿色荧光,胞核为蓝色。结论磷酸化组蛋白H3是观察和鉴别心肌细胞有丝分裂的可靠指标。Objective Applying a reliable precise method to assess the mitotic index of cardiomyocytes,to disclose of disclosure the mechanism implicated in cardiomyocytes proliferation. Methods H9c2（2-1） cardiomyocytes were originally developed from rat BD1X heart（ATCC）.These cells were cultured on coverslips.Double immunofluorescence staining with monoclonal antibodies（1：100） against phospho-histone H3 and α-sarcomeric actin was performed on the cultured cells.Anti-mouse IgG FITC was used as the secondary antibody for the H3P antibody,and anti-mouse IgM Cy3 was used as the secondary antibody for the α-sarcomeric actin.DNA was visualized with Hochest 33342.All photographs were taken with an Olympus fluorescence microscope. Results The cytoplasm of cardiomyocytes appeared red,the mitotic chromosomes green with distinct shape,and Hochest 33342-stained nuclei blue.Conclusion Our method is the reliable and exact means to observe and assess cardiomyocytes mitosis.

关 键 词：H9c2(2—1)细胞 有丝分裂 指数 磷酸化组蛋白H3 横纹肌肌动蛋白 免疫荧光双重染色 大鼠 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]