表皮细胞去分化的体内试验研究.DOCVIP

在体诱导表皮细胞去分化的实验研究 蔡飒 付小兵郭永峰 孙同柱 王君 李海红 （中国人民解放军总医院第一附属医院全军创伤修复重点实验室目的 在体诱导表皮细胞去分化，探索与表皮细胞去分化有关的信号通路。方法 包皮皮片去除皮下组织，用中性蛋白酶分离表皮，Ⅳ型胶原反复粘贴表皮去除基底细胞层。处理后表皮片移植于裸鼠全层皮肤下，分别于3，5，7d取出移植皮片，采用免疫组织化学染色，Western blot和RT-PCR等方法检测皮片移植前后表型的改变和ERK MAPK通路各信号分子表达的变化情况。结果 表皮片经去基底层处理后，表皮角消失，CK19，(1整合素，Oct4和p63染色阴性。移植后大部分皮片柔软、红润，存活率为86.3%。移植后存活皮片CK19，(1整合素，Oct4和p63染色阳性细胞分散分布于基底上层，CK10阳性细胞数量减少；CK19，(1整合素，Oct4和p63等指标蛋白及mRNA表达增强，其中以移植5d组最为明显；ERK及上下游信号分子（p- Raf、p-MEK1/2、p-ERK1/2、和c-myc）表达增强。t-MEK1/2和t-ERK1/2移植前后表达无显著差异。结论 处于终末分化阶段的表皮细胞可以向干细胞状态逆转，这个过程可能与ERK MAPK信号通路有关。 关键词：去分化；异种移植；表皮细胞；表皮干细胞；胞外信号调节激酶 基金项目：国家重大基础研究规划资助项目（2005 CB 522603） 国家自然科学基金资助项目（2005 作者：cai_sa@ 通讯作者：付小兵E-mail: fuxiaobing@ The in vivo study of dedifferentiation of epidermal cells CAI Sa，FU Xiao-bing，GUO Yong-feng，SUN Tong-zhu，WANG Jun，LI Hai-hong Wound Healing and Cell Biology Laboratory Burns Institute, The First Affiliated Hospital (304th Hospital) General Hospital of PLA, Trauma Center of Postgraduate Medical College, Beijing 100037, China Corresponding author: Fu Xiaobing, E-mail: fuxb@ fuxiaobing@ Abstract：Objective to induce dedifferentiation of epidermal cells in vivo and explore the signaling pathway involved in the process. Methods the epidermis of human foreskin was isolated from dermis by digestion with protease, and the cells in the stratum basale of the epidermis together with stem cells were eliminated by repeated adhesion to collagen type Ⅳ. After being labeled with DAPI, the treated ultrathin epidermal sheets were transplanted under the fresh full-thickness skin wounds measuring about 1 cm in diameter on the back of athymic BALB /c nude mice. The transplanted ultrathin epidermal sheets were harvested from the recipient nude mice at d 3, 5 and 7 after xenotransplantation. Immunohistochemical, Western blot and RT-PCR analyses were used to determine the changes in phenotype and in ERK signaling pathway of the xenografts. Results CK19, (1 integrin, p63 and Oct4 p