不同发育阶段人皮肤表皮干细胞增殖分化相关基因的表达特征  

Expression characteristics of human skin epidermal stem cell proliferation and differentiation-related genes during different developmental periods

作　　者：黄培信[1] 刘德伍[2] 王肖蓉[1] 毛远桂 陈剑平[2] 蓝蔚[2] 

机构地区：[1]河南科技大学第一附属医院烧伤整形科,河南省洛阳市471003 [2]南昌大学第一附属医院烧伤研究所,江西省南昌市330006

出　　处：《中国组织工程研究与临床康复》2009年第23期4442-4446,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30560058);江西省科技厅重大科技招标项目(200604)~~

摘　　要：背景：目前研究认为表皮干细胞数量及增殖分化潜能的差异，可能是创面愈合能力随年龄增大而逐渐降低的重要原因之一。目的：探讨不同发育阶段人皮肤表皮干细胞B1整合素、角蛋白19、p63转录因子和端粒酶反转录酶的表达特征。设计、时间及地点：细胞学体外对照观察，于2005—09／2007—04在南昌大学第一附属医院烧伤研究所完成。材料：流产胎儿皮肤标本由南昌大学第一附属医院产科提供，4～12周岁少儿、25～45岁成人烧伤整形植皮手术剩余皮片标本由南昌大学第～附属医院烧伤科提供。方法：标本取材后立即用中性甲醛固定，常规石蜡包埋，4℃保存。所有标本经系列切片后，分别采用鼠抗人角蛋白19、β1整合素、p63转录因子、端粒酶反转录酶单克隆抗体作为一抗，以免疫组织化学EnVision法检测皮肤组织中表皮干细胞标志物的表达。主要观察指标：200倍光镜下观察免疫组化染色结果，测定表皮干细胞标志物阳性表达率。结果：胎儿皮肤、少儿皮肤和成人皮肤表皮的基底部、毛囊周围细胞B1整合素、角蛋白19、p63和端粒酶反转录酶均呈棕黄色阳性表达。胎儿表皮基底层细胞β1整合素、角蛋白19和p63均呈强阳性表达：少儿表皮基底层细胞叶1大部分表达β1整合素、角蛋白19和p63，阳性细胞均匀分布；成人表皮基底层细胞B1整合素、角蛋白19和p63呈弱阳性表达，阳性细胞散在分布；胎儿表皮基底层端粒酶反转录酶阳性细胞表达强度，少儿表皮〉成人表皮。随年龄的增加，β1整合素、角蛋白19、p63和端粒酶反转录酶在胎儿、少儿、成人皮肤组织中的阳性表达率呈下降趋势（F=5．06，P〈0．01）。结论：胎儿皮肤、少儿皮肤和成人皮肤表皮干细胞主要位于表皮基底层，端粒酶表达阳性区域及信号强度与表皮干细胞的定位分布及阳性表达强度相�BACKGROUND： The difference in number, proliferation and differentiation of epidermal stem cells may be a key reason for the decrease in wound healing ability with age. OBJECTIVE： To study the expression characteristics of 131 integrin, keratin-19, p63 transcriptional factor and telomerase reverse transcriptase during different developmental periods of human epidermal stem cells. DESIGN, TIME AND SETTING： The cytological in vitro controlled study was performed at the Institute of Burn, First Affiliated Hospital, Nanchang University from September 2005 to April 2007. MATERIALS： Skin samples of aborted fetus were supplied by the Department of Obstetrics, First Affiliated Hospital, Nanchang University. Remaining skin samples of 4 12 children and 25-45 adults following surgery were obtained from the Department of Burn, First Affiliated Hospital, Nanchang University. METHODS： Following sampling, these samples were fixed in neutral formalin, embeded into paraffin, placed at 4℃. After series of sections, integrin β1, keratin19, p63 transcription factor and telomerase reverse transcriptase monoclonal antibody were considered as the primary antibody. Immunohistochemical EnVision method was used to detect expression of epidermal stem cell markers in skin tissues. MAIN OUTCOME MEASURES： Optical microscope at 200x magnification was used to observe the staining results. Positive expression rate of epidermal stem cel markers was measured. RESULTS： The β1 integrin, keratin 19, p63 and telomerase reverse transcriptase positive cells were mainly located in the epidermal basement membrane, and the expression levels of those indexes were highest in the fetus skin, lower in the skins of the child and adult, the distribution was uniformity in the fetus and adult samples. Lowest expression was found in the adult sample. The expression of β1 integrin, keratin 19, p63 and telomerase reverse transcriptase in the different development periods revealed decreasing tendency with the age increasing （F = 5.06, P 〈 0

关 键 词：表皮干细胞 皮肤 端粒酶 创面修复 

分 类 号：R394.2[医药卫生—医学遗传学]