Wnt10b诱导再生毛囊的表达特性研究

中华皮肤科杂志 ›› 2016, Vol. 49 ›› Issue (4): 261-264.

Wnt10b诱导再生毛囊的表达特性研究

星懿展,郭海英,马小艮,李玉红

中国人民解放军第三军医大学

收稿日期:2015-09-25 修回日期:2015-10-31 出版日期:2016-04-15 发布日期:2016-03-28
通讯作者: 李玉红 E-mail:liyuhongtmmu@hotmail.com
基金资助:
Wnt10b-BMP6间的活化-抑制信号交流调控毛囊周期性再生的研究;Wnt10b 激活毛囊干细胞的作用与机理研

Expression characteristics in regenerating hair follicles induced by Wnt10b

Received:2015-09-25 Revised:2015-10-31 Online:2016-04-15 Published:2016-03-28
Contact: Yu-Hong LI E-mail:liyuhongtmmu@hotmail.com

摘要/Abstract

摘要：

目的研究Wnt10b诱导再生毛囊的表达特性及诱导作用机制。方法 HEK-293细胞内扩增并用氯化铯梯度离心纯化Wnt10b过表达腺病毒及对照腺病毒，皮内注射至C57BL/6J小鼠背部皮肤，在处理后2.5、5、7、9、14、28 d时取材，HE染色及免疫组化染色观察毛囊结构特征、信号通路表达特征及增殖特性。结果 HE染色发现，AdWnt10b处理组从第5天开始出现新生毛囊结构，正常生长，第28天左右进入退化期。免疫组化染色发现，AdWnt10b处理组从处理后5 d开始新生毛囊具有AE15表达，随着毛囊生长而增加，至处理后28 d开始减少。在AdWnt10b处理后5 d，观察到β连环素的核表达，Lef1特异性表达于毛芽和毛母质部位，且全为核表达。在AdWnt10b处理后28 d，Lef1表达减弱。AdWnt10b处理后2.5 d即可见Ki67表达于表皮和毛囊外根鞘。处理后2.5、7、9、14 d均在隆突区见到Ki67的表达；从处理后7 d开始，Ki67表达于毛母质细胞。结论 Wnt10b诱导的再生毛囊具有正常的毛囊结构，Wnt10b激活了经典Wnt信号通路，其作用的靶细胞是毛囊干细胞及其子代细胞。

关键词: &beta, 连环素

Abstract:

Xing Yizhan, Guo Haiying, Ma Xiaogen, Li Yuhong Department of Cell Biology, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China Corresponding author: Li Yuhong, Email: liyuhongtmmu@hotmail.com 【Abstract】 Objective To investigate characteristics in regenerating hair follicles induced by Wnt10b, and to explore mechanisms underlying Wnt10b-induced regeneration of hair follicles. Methods Both adenovirus containing the Wnt10b gene （AdWnt10b） and that containing the green fluorescent protein-encoding gene （AdGFP） were amplified in HEK-293 cells and purified by caesium chloride density gradient centrifugation. A total of 36 C57BL/6J mice were randomly and equally divided into the AdWnt10b group and AdGFP group to be intracutaneously injected with AdWnt10b and AdGFP on the back respectively. Three mice were sacrificed on day 2.5, 5, 7, 9, 14 and 28 after the injection separately, and skin samples were resected from the injected sites subsequently. Hematoxylin and eosin （HE） staining and immunohistochemical staining were performed to observe hair follicle structure, analyze characteristics of the Wnt signaling pathway, and to estimate proliferative activity of regenerating hair follicles induced by Wnt10b. Results As HE staining showed, new hair follicles appeared as early as day 5 after the injection, then grew normally, and entered into catagen phase on day 28 in the AdWnt10b group. Immunohistochemical staining showed that AE15 was observed in new hair follicles as early as day 5 after the injection in the AdWnt10b group, then increased along with the growth of hair follicles, but decreased on day 28. On day 5 after AdWnt10b injection, both β-catenin and Lef1 s were seen in the cell nucleus. Lef1 was expressed specifically in hair germs and hair matrix, and its began to decrease on day 28. In addition, Ki67 was observed in the epidermis and outer root sheath of hair follicles as early as day 2.5 after the injection, in the bulge region of hair follicles on day 2.5, 7, 9 and 14, and in hair matrix cells as early as day 7. Conclusion Wnt10b could induce regeneration of hair follicles with normal structure, likely by activating the canonical Wnt signaling pathway in hair follicle stem cells and their daughter cells.

星懿展郭海英马小艮李玉红. Wnt10b诱导再生毛囊的表达特性研究[J]. 中华皮肤科杂志, 2016, 49(4): 261-264.

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