大鼠生后不同发育阶段心肌细胞有丝分裂指数检测  

Detection of Mitotic Index of Cardiomyocytes during the Terminal Differentiation of the Postnatal Rat Heart

作　　者：邸菁[1] 陈双叶[2] 赵晨超[2] 张凌志[3] 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,辽宁沈阳110001 [2]中国医科大学,89期七年制辽宁沈阳110001 [3]中国医科大学附属盛京医院急诊科,辽宁沈阳110004

出　　处：《解剖科学进展》2007年第3期203-206,共4页Progress of Anatomical Sciences

基　　金：辽宁省教育厅资助项目(No.051525)

摘　　要：目的检测大鼠生后不同发育阶段心肌细胞的有丝分裂指数,为研究心肌细胞增殖机制奠定形态学基础。方法取大鼠生后不同发育阶段心肌组织行冰冻切片,采用免疫荧光双重染色方法,以横纹肌肌动蛋白单克隆抗体标记心肌细胞,以磷酸化组蛋白H3单克隆抗体标记有丝分裂的染色体,同时用Hochest33342显示细胞核,荧光显微镜观察并拍照。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;大鼠生后零天心肌细胞有丝分裂指数(0.905±0.087%)是生后2w(0.372±0.094%)的2.4倍(P<0.01),4w至成年几乎为零。结论大鼠出生2w后心肌细胞逐渐丧失增殖能力,由增生性生长向肥大性生长转化。Objective To assess the mitotic index of cardiomyocytes during the terminal differentiation of the postnatal rat heart, for the purpose of disclosure the mechanism implicated in cardiomyocytes proliferation. Methods Whole ventricular frozen sections from postnatal（PN0, PN14, PN28） and adult hearts were fixed in 4% paraformaldehyde. Double immunofluorescence staining of monoclonal antibodies （1：100） against phospho-histone H3 （ H3P ） and α - sareomeric actin was performed on the frozen sections. Anti-mouse IgG FITC was used as the secondary antibody for the H3P antibody, and anti-mouse IgM Cy3 for the α -sarcomeric actin. DNA was visualized with Hochest33342. All photographs were taken with an olympus fluorescence microscope. Then mitotic index of cardiomyocytes was calculated. Results The cytoplasm of the cardiomyocytes appeared red, the mitotic chromosomes appeard green with distinct shape, and Hochest33342-stained nuclei appeard blue. The mitotic index of cardiomyocytes in 0-day-old hearts （0.905 ± 0.087%） was approximately 2.4 times over that in 2-week-old hearts（0.372 ± 0.094%）. Conclusion The majority of cardiomyocytes in the rat heart lost the ability of differentiation and proliferation two weeks after birth.

关 键 词：大鼠 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 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]