中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (7): 1343-1349.doi: 10.12307/2025.023

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

过表达神经调节蛋白1的人羊膜间充质干细胞促进小鼠皮肤创面愈合

胡涛涛，刘  兵，陈  诚，殷宗银，阚道洪，倪  杰，叶凌霄，郑祥兵，严  敏，邹  勇   

  1. 四川大学华西医院宜宾医院，四川省宜宾市   644000
  • 收稿日期:2023-10-17 接受日期:2024-02-25 出版日期:2025-03-08 发布日期:2024-06-27
  • 通讯作者: 邹勇，硕士，主任医师，四川大学华西医院宜宾医院，四川省宜宾市 644000
  • 作者简介:胡涛涛，男，1992年生，四川省渠县人，汉族，2022年遵义医科大学毕业，硕士，主要从事创面修复与神经再生相关研究。
  • 基金资助:
    宜宾市第二人民医院院级孵育项目(2021FY16)，项目负责人：阚道洪

Human amniotic mesenchymal stem cells overexpressing neuregulin-1 promote skin wound healing in mice

Hu Taotao, Liu Bing, Chen Cheng, Yin Zongyin, Kan Daohong, Ni Jie, Ye Lingxiao, Zheng Xiangbing, Yan Min, Zou Yong   

  1. Yibin Hospital, West China Hospital, Sichuan University, Yibin 644000, Sichuan Province, China
  • Received:2023-10-17 Accepted:2024-02-25 Online:2025-03-08 Published:2024-06-27
  • Contact: Zou Yong, Master, Chief physician, Yibin Hospital, West China Hospital, Sichuan University, Yibin 644000, Sichuan Province, China
  • About author:Hu Taotao, Master, Yibin Hospital, West China Hospital, Sichuan University, Yibin 644000, Sichuan Province, China
  • Supported by:
    Yibin Second People’s Hospital Hospital-level Incubation Program, No. 2021FY16 (to KDH)

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摘要：

文题释义：

人羊膜间充质干细胞：来源于人胎盘羊膜组织的间充质干细胞，具有多向分化潜能，能合成并分泌各种细胞外基质蛋白、细胞因子、生长因子以及其他有助于愈合和再生的生物活性蛋白，目前已被广泛用于组织修复相关研究，是组织工程领域重要的种子细胞。
神经调节蛋白1：是一类与表皮生长因子密切相关的生长与分化因子家族。神经调节蛋白1受体是酪氨酸激酶转膜受体家族，广泛参与细胞的增殖、分化、迁移、生长、存活以及凋亡。


背景：神经调节蛋白1具有促进细胞增殖、分化以及血管生长等特性。人羊膜间充质干细胞是组织工程领域重要的种子细胞，已被证实参与组织修复及再生过程。
目的：构建过表达神经调节蛋白1的人羊膜间充质干细胞，探究其增殖、迁移能力以及对创面愈合的影响。
方法：①体外分离培养人羊膜间充质干细胞并对其进行鉴定；②构建神经调节蛋白1过表达慢病毒，将人羊膜间充质干细胞分为空载组、神经调节蛋白1组、对照组，分别转染空载慢病毒、过表达神经调节蛋白1慢病毒，对照组不进行转染；③EdU实验检测各组细胞增殖能力，Transwell实验检测各组细胞迁移能力；④构建C57BL/6小鼠创面损伤模型，随机分成对照组、空载组和神经调节蛋白1组，每组8只，分别在创面局部多点均匀注射1 mL转染空载慢病毒或转染过表达神经调节蛋白1慢病毒的人羊膜间充质干细胞，对照组注射等量的生理盐水；⑤造模后1，7，14 d观察创面愈合情况，苏木精-伊红染色观察创面愈合组织学变化，免疫组化观察创面CD31的表达。
结果与结论：①成功构建过表达神经调节蛋白1的人羊膜间充质干细胞，细胞内神经调节蛋白1的mRNA、蛋白表达较空载组明显上调(P < 0.05)；②过表达神经调节蛋白1促进了人羊膜间充质干细胞的迁移(P < 0.01)和增殖(P < 0.05)；③过表达神经调节蛋白1的人羊膜间充质干细胞促进了小鼠创面愈合(P < 0.05)和创面的血管生成(P < 0.05)。结果表明，过表达神经调节蛋白1提高了人羊膜间充质干细胞的增殖和迁移能力，以及增强了促进创面愈合和创面血管生成的能力。

https://orcid.org/0000-0003-0655-2810 (胡涛涛) 


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: 人羊膜间充质干细胞, 神经调节蛋白1, 过表达, 细胞增殖, 细胞迁移, 小鼠, 创面, 愈合

Abstract: BACKGROUND: Neuregulin 1 has been shown to be characterized in cell proliferation, differentiation, and vascular growth. Human amniotic mesenchymal stem cells are important seed cells in the field of tissue engineering, and have been shown to be involved in tissue repair and regeneration.
OBJECTIVE: To construct human amniotic mesenchymal stem cells overexpressing neuregulin 1 and investigate their proliferation and migration abilities, as well as their effects on wound healing.
METHODS: (1) Human amniotic mesenchymal stem cells were in vitro isolated and cultured and identified. (2) A lentivirus overexpressing neuregulin 1 was constructed. Human amniotic mesenchymal stem cells were divided into empty group, neuregulin 1 group, and control group, and transfected with empty lentivirus and lentivirus overexpressing neuregulin 1, or not transfected, respectively. (3) Edu assay was used to detect the proliferation ability of the cells of each group, and Transwell assay was used to detect the migration ability of the cells. (4) The C57 BL/6 mouse trauma models were constructed and randomly divided into control group, empty group, neuregulin 1 group, with 8 mice in each group. Human amniotic mesenchymal stem cells transfected with empty lentivirus or lentivirus overexpressing neuregulin-1 were uniformly injected with 1 mL at multiple local wound sites. The control group was injected with an equal amount of saline. (5) The healing of the trauma was observed at 1, 7, and 14 days after model establishment. Histological changes of the healing of the trauma were observed by hematoxylin-eosin staining. The expression of CD31 on the trauma was observed by immunohistochemistry. 
RESULTS AND CONCLUSION: (1) Human amniotic mesenchymal stem cells overexpressing neuregulin-1 were successfully constructed. The mRNA and protein expression of intracellular neuregulin 1 was significantly up-regulated compared with the empty group (P < 0.05). (2) The overexpression of neuregulin 1 promoted the migratory ability (P < 0.01) and proliferative ability of human amniotic mesenchymal stem cells (P < 0.05). (3) Human amniotic mesenchymal stem cells overexpressing neuregulin 1 promoted wound healing in mice (P < 0.05) and wound angiogenesis (P < 0.05). The results showed that overexpression of neuregulin 1 resulted in an increase in the proliferative and migratory capacities of human amniotic mesenchymal stem cells, significantly promoting wound healing and angiogenesis.

Key words: human amniotic mesenchymal stem cell, neuregulin-1, overexpression, cell proliferation, cell migration, mouse, wound, healing ,

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