中国组织工程研究 ›› 2020, Vol. 24 ›› Issue (31): 5018-5022.doi: 10.3969/j.issn.2095-4344.2136

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

鼠胚胎期和出生后毛囊下段启动再生表达的Wnt5a

蔡博治1,陈先才2,倪  娜3,林伟深4,袁燕平5,林常敏6,黄  铿4   

  1. 1汕头大学医学院第一附属医院分子心脏病学实验室,广东省汕头市  515041;2广西贵港市贵港华美医疗美容门诊部,广西壮族自治区贵港市  537100;汕头大学医学院,3临床技能中心,6组织与胚胎学教研室,广东省汕头市  515041;4汕头大学医学院第二附属医院急诊科,广东省汕头市  515041;5江西省九江学院附属医院病理科,江西省九江市  332000
  • 收稿日期:2020-02-12 修回日期:2020-02-21 接受日期:2020-03-24 出版日期:2020-11-08 发布日期:2020-09-04
  • 通讯作者: 黄铿,主任医师,硕士生导师,汕头大学医学院第二附属医院急诊科,广东省汕头市 515041
  • 作者简介:蔡博治,男,1987年生,广东省汕头市人,汉族,2014年汕头大学毕业,助理实验师,主要从事组织工程化毛囊研究。
  • 基金资助:
    国家自然科学基金(81972957);广东省科技发展专项资金(2017A030313776)

Wnt5a expression in the regeneration of the inferior segment of rat hair follicle at embryonic and postnatal stages 

Cai Bozhi1, Chen Xiancai2, Ni Na3, Lin Weishen4, Yuan Yanping5, Lin Changmin6, Huang Keng4   

  • Received:2020-02-12 Revised:2020-02-21 Accepted:2020-03-24 Online:2020-11-08 Published:2020-09-04
  • Contact: Huang Keng, Chief physician, Master’s supervisor, Emergency Department of the Second Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong Province, China
  • About author:Cai Bozhi, Assistant experimentalist, Molecular Cardiology Laboratory of the First Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong Province, China
  • Supported by:
    the National Natural Science Foundation of China No. 81972957 ; the Specific Scientific Development Foundation of Guangdong Province, No. 2017A030313776 

摘要:

文题释义:

毛囊下段再生从大鼠触须垫分离出完整的触须毛囊,在镜下紧贴毛球部上方切除毛球部并拔除毛干,将毛囊上段移植到裸鼠皮下,观察毛囊下段再生的情况。

Wnt5aWnt蛋白家族中的成员之一,在细胞成熟、胚胎发育等过程中发挥着重要作用。研究表明Wnt10aWnt5a的表达在毛囊发生早期(E14.5)显著地上调,其中Wnt5a是唯一在真皮高表达的Wnt信号。

背景:Wnt5a是唯一在真皮高表达,与毛囊真皮成分发生密切相关的信号分子。

目的:通过观察E12.5-E15.5胎龄的大鼠胚胎皮肤和大鼠触须毛囊上段裸鼠移植诱导毛乳头再生过程Wnt5a的表达,初步研究毛乳头胚胎发生和体外再生的启动机制。

方法:将孕SD大鼠在E12.5E13.5E14.5E15.5麻醉后处死,取出完整胚胎制作石蜡切片;另取单个SD大鼠触须毛囊切除毛乳头,毛囊上段拔除毛干后移植至Nu/Nu裸鼠背部皮肤;分别取以上胎龄的大鼠胚胎皮肤及裸鼠背部移植部位的毛囊标本,用酶标免疫组化法观察Wnt5a在大鼠胚胎皮肤及大鼠触须毛囊上段裸鼠移植诱导毛乳头再生过程的表达,苏木精-伊红染色观察大鼠触须毛囊上段移植后毛囊下段的再生情况。动物实验均已获得汕头大学医学院动物伦理委员会批准。

结果与结论:①E12.5 Wnt5a在表皮细胞表达;E13.5 Wnt5a在表皮细胞表达增强;E14.5 Wnt5a表达由表皮转移至真皮,在真皮细胞浆明显表达;E15.5 Wnt5a表达在真皮结缔组织明显增强,在表皮的表达消失;②大鼠触须毛囊上段移植后第3天开始至第9天,毛囊上段向下形成新的毛球部,恢复完整毛囊结构,再生过程相关组织Wnt5a均呈强阳性表达;③Wnt5a在胚胎毛囊发生前后和毛囊下段再生模型毛乳头形成的部位呈现高表达,说明Wnt信号通路与毛乳头再生有密切的关系,其中Wnt5a可能扮演了非常主要的角色。

ORCID: 0000-0003-4428-6680(蔡博治)

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关键词: 毛囊, 毛乳头,  真皮,  胚胎;,  通路,  鼠,  实验,  再生

Abstract:

BACKGROUND: Wnt5a is the only signaling molecule that is highly expressed in the dermis and closely related to the occurrence of hair follicle dermal components.

OBJECTIVE: To preliminarily discuss initiation mechanism of dermal papilla embryogenesis and regeneration in vitro by observing Wnt5a expression of E12.5-E15.5 fetal rat skin and the upper part of tentacle hair follicle during dermal papilla regeneration induced by transplantation.

METHODS: The pregnant Sprague-Dawley rats at E12.5, E13.5, E14.5 and E15.5 days were killed under anesthesia. The embryos were taken out to make paraffin sections. The hairy papilla was removed from the tentacle hair follicles of Sprague-Dawley rats, and the hair follicles were removed from the upper segment of the hair follicles and then injected into the skin of the back of Nu/Nu nude mice. The skins of rat embryos at above gestational age and hair follicles at transplanted back positions of the nude mice were taken. Enzymatic immunohistochemical method was used to observe the Wnt5a expression in rat embryonic skin and the upper segment of hair follicle during dermal papilla regeneration induced by transplantation. Hematoxylin-eosin staining was used to observe the regeneration of the inferior segment of hair follicle after transplantation in the upper segment. The animal experiment was approved by the Animal Ethics Committee of Shantou University Medical College.

RESULTS AND CONCLUSION: (1) E12.5 Wnt5a was expressed in epidermal cells; E13.5 Wnt5a was expressed in epidermal cells and enhanced; E14.5 Wnt5a was transferred from epidermis to dermis and expressed in dermal cytoplasm; E15.5 Wnt5a was expressed in dermal connective tissue, obviously enhanced, and disappeared in epidermis. (2) From the third day to the ninth day after transplantation of the upper part of the rat hair follicle, the upper fragment of the hair follicle was observed to be growing downward. The complete follicles were re-formed. Wnt 5A was strongly expressed in the regeneration related tissues. (3) The high expression of Wnt5a in the site of dermal papilla formation in this model and dermal papilla embryogenesis suggests that Wnt signaling pathway is closely related to the regeneration of dermal papilla, and Wnt5a may play a very important role. 

Key words: hair follicle,  dermal papilla,  dermis,  embryo,  pathway,  mouse,  experiment,  regeneration

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