[1]蔡飒,付小兵,雷永红,等.在体诱导表皮细胞去分化的初步实验研究[J].第三军医大学学报,2008,30(21):1998-2002.
 CAI Sa,FU Xiao-bing,LEI Yong-hong,et al.Dedifferentiation of human epidermal cells in nude mice[J].J Third Mil Med Univ,2008,30(21):1998-2002.
点击复制

在体诱导表皮细胞去分化的初步实验研究(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第21期
页码:
1998-2002
栏目:
论著
出版日期:
2008-11-15

文章信息/Info

Title:
Dedifferentiation of human epidermal cells in nude mice
作者:
蔡飒付小兵雷永红韩冰郭永峰孙同柱王君李海红
解放军总医院第一附属医院全军创伤修复重点实验室
Author(s):
CAI Sa FU Xiao-bing LEI Yong-hong HAN Bing GUO Yong-feng SUN Tong-zhu WANG Jun LI Hai-hong
Wound Healing and Cell Biology Laboratory, Burns Institute, First Affiliated Hospital,  General Hospital of PLA, Beijing 100037, China
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
Keywords:
dedifferentiation xenotransplantation epidermal cells epidermal stem cells extracellular signal regulated kinase
分类号:
R-332; R322.991; R329.2
文献标志码:
A
摘要:
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型胶原反复粘贴并牵拉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、7 d取出移植皮片,采用免疫组织化学染色、流式细胞仪检测、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    去基底细胞层处理的表皮片干细胞标志物CK19和β1整合素染色阴性;移植后5 d,存活皮片CK19和β1整合素染色呈多层分布,而不是正常表皮中的单层分布。流式细胞仪检测结果显示,表皮片移植后CK19阳性细胞和β1整合素阳性细胞由移植前的0.00%分别增加到7.54%和5.24%,而CK10阳性细胞由99.62%下降为86.56%。Western blot和PCR检测也证实CK19和β1整合素蛋白水平和mRNA水平表达在移植后明显增强。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(P>0.05)。    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。
Abstract:
Objective    To induce the dedifferentiation of human epidermal cells in vivo for the signaling pathways involved in the process.     Methods    The epidermis of human foreskin which was obtained with informed consent and approval of the Ethics Committee of our hospital was isolated from dermis by digestion with protease, and the cells in the stratum basale of the epidermis as well as 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 about 1 cm in diameter on the back of athymic BALB/c nude mice. The transplanted sheets were harvested from the recipient at day 3, 5 and 7 after xenotransplantation. Immunohistochemical assay, Western blot analysis and RT-PCR were used to determine the changes of cell phenotypes and ERK signaling pathway of the xenografts.     Results    CK19, β1 integrin, p63 and Oct4 positive cells were found in a single layer in stratum basal of the full-thickness epidermis, but were undetectable in ultrathin epidermal sheets before grafting. Most of the ultrathin epidermal sheets grew well in a soft and ruddy appearance after transplantation and the total survival rate was 86.3%. Immunological examinations showed that a certain number of cells were positive to CK19, β1 integrin, p63 and Oct4 in spinous and granular layers after transplantation and that ERK/MAPK pathway was involved in the reversion process.     Conclusion    The differentiating epidermal cells could reverse to stem cell-like cells, and the process might be associated with the regulation of ERK/MAPK signaling pathway.

参考文献/References:

蔡飒, 付小兵, 雷永红, 等. 在体诱导表皮细胞去分化的初步实验研究[J]. 第三军医大学学报, 2008, 30(21):1998-2002.

相似文献/References:

[1]杨光伦,姚榛祥,黄平,等.来氟米特在大鼠异种心脏移植排斥反应中对NF-κB P65的影响[J].第三军医大学学报,2007,29(23):2267.
 YANG Guang-lun,YAO Zhen-xiang,HUANG Ping,et al.Effect of leflunomide on NF-κB P65 in mouse to rat cardiac xenograft[J].J Third Mil Med Univ,2007,29(21):2267.
[2]杨光伦,姚榛祥,黄平,等.NF-κB P65在大鼠异种心脏移植后心肌中的表达及意义[J].第三军医大学学报,2007,29(01):51.
 YANG Guang-lun,YAO Zhen-xiang,HUANG Ping,et al.Expression of NF-κB P65 in mouse to rat cardiac xenograft model[J].J Third Mil Med Univ,2007,29(21):51.
[3]张翠萍,陈鹏,付小兵,等.Wnt/β-catenin信号通路活化对人表皮细胞表型变化的影响[J].第三军医大学学报,2010,32(16):1716.
 Zhang Cuiping,Chen Peng,Fu Xiaobing,et al.Effect of Wnt/β-catenin pathway activation on phenotypic characteristics of epidermal cells in vitro[J].J Third Mil Med Univ,2010,32(21):1716.
[4]黄正根,吴军,罗高兴,等.胎猪皮肤前体组织异种移植模型的研究[J].第三军医大学学报,2008,30(22):2072.
 HUANG Zheng-gen,WU Jun,LUO Gao-xing,et al.Xenotransplantation model of fetal pig skin precursor tissue[J].J Third Mil Med Univ,2008,30(21):2072.
[5]孙晓艳,付小兵,伊海英,等.碱性成纤维细胞生长因子诱导表皮细胞去分化的初步研究[J].第三军医大学学报,2008,30(21):2003.
 SUN Xiao-yan,FU Xiao-bing,YI Hai-ying,et al.bFGF-induced dedifferentiation of epidermal cells into epidermal progenitor stem cells[J].J Third Mil Med Univ,2008,30(21):2003.
[6]唐华,姚榛祥,刘盛春.协调性异种心脏移植排斥反应病理变化的动态研究[J].第三军医大学学报,2007,29(08):721.
 TANG Hua,YAO Zhen-xiang,LIU Sheng-chun.Dynamic pathological changes in concordant cardiac xenograft rejection[J].J Third Mil Med Univ,2007,29(21):721.
[7]黄辉煌,杨列,陈卫军,等.中国版纳小型猪近交系肝脏的应用解剖[J].第三军医大学学报,2004,26(03):0.[doi:10.16016/j.1000-5404.2004.03.012 ]
 HUANG Hui huang,YANG Lie,CHEN Wei jun,et al.[J].J Third Mil Med Univ,2004,26(21):0.[doi:10.16016/j.1000-5404.2004.03.012 ]
[8]徐承平,卞修武,章容,等.去甲二氢愈创木酸对人恶性胶质瘤细胞系CHG-5生长的影响[J].第三军医大学学报,2004,26(08):0.[doi:10.16016/j.1000-5404.2004.08.006 ]
 XU Cheng ping,BIAN Xiu wu,ZHANG Rong,et al.[J].J Third Mil Med Univ,2004,26(21):0.[doi:10.16016/j.1000-5404.2004.08.006 ]
[9]丁生财,陈意生,魏泓,等.猪皮肤成纤维细胞的培养与鉴定[J].第三军医大学学报,2004,26(13):0.[doi:10.16016/j.1000-5404.2004.13.017 ]
 DING Sheng-cai,CHEN-Yi-sheng,WEI Hong,et al.[J].J Third Mil Med Univ,2004,26(21):0.[doi:10.16016/j.1000-5404.2004.13.017 ]
[10]贺伟峰,吴军,罗高兴,等.大鼠树突状细胞的体外分离、纯化、扩增及鉴定[J].第三军医大学学报,2002,24(08):0.[doi:10.16016/j.1000-5404.2002.08.001 ]
 HE Wei-feng,WU Jun,LUO Gao xing,et al.[J].J Third Mil Med Univ,2002,24(21):0.[doi:10.16016/j.1000-5404.2002.08.001 ]

更新日期/Last Update: 2008-10-20