中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (33): 5249-5255.doi: 10.12307/2023.750

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

毛囊表皮神经嵴干细胞调节面神经损伤后局部炎症因子的表达水平

唐  笠，潘  瑶，朱国臣   

  1. 南京医科大学附属无锡第二医院耳鼻咽喉头颈外科，江苏省无锡市   214002
  • 收稿日期:2022-10-24 接受日期:2022-12-09 出版日期:2023-11-28 发布日期:2023-03-29
  • 通讯作者: 朱国臣，博士，主任医师，教授，硕士生导师，南京医科大学附属无锡第二医院耳鼻咽喉头颈外科，江苏省无锡市 214002
  • 作者简介:唐笠，男，1996年生，浙江省金华市人，汉族，南京医科大学在读硕士，主要从事面神经损伤与修复的基础及临床工作。
  • 基金资助:
    国家自然科学基金面上项目(81770990)，项目负责人：朱国臣；江苏省科技厅社会发展重点研发项目(BE2018628)，项目负责人：朱国臣；江苏省第十六批“六大人才高峰”高层次人才(WSW-141)，项目负责人：朱国臣；无锡市科技局-医疗卫生技术攻关项目(Y20212003)，项目负责人：朱国臣；无锡市卫健委精准医学项目(J202002)，项目负责人：朱国臣

Epidermal neural crest stem cells of hair follicles regulate the local expression of inflammatory factors after facial nerve injury

Tang Li, Pan Yao, Zhu Guochen   

  1. Department of Otorhinolaryngology-Head and Neck Surgery, Affiliated Wuxi No. 2 People’s Hospital, Nanjing Medical University, Wuxi 214002, Jiangsu Province, China
  • Received:2022-10-24 Accepted:2022-12-09 Online:2023-11-28 Published:2023-03-29
  • Contact: Zhu Guochen, MD, Chief physician, Professor, Master’s supervisor, Department of Otorhinolaryngology-Head and Neck Surgery, Affiliated Wuxi No. 2 People’s Hospital, Nanjing Medical University, Wuxi 214002, Jiangsu Province, China
  • About author:Tang Li, Master candidate, Department of Otorhinolaryngology-Head and Neck Surgery, Affiliated Wuxi No. 2 People’s Hospital, Nanjing Medical University, Wuxi 214002, Jiangsu Province, China
  • Supported by:
    General Program of the National Natural Science Foundation of China, No. 81770990 (to ZGC); Social Development Key R&D Project of Jiangsu Provincial Science and Technology Department, No. BE2018628 (to ZGC); The 16th Batch of “Six Talent Peaks” High-Level Talents in Jiangsu Province, No. WSW-141 (to ZGC); Wuxi Science and Technology Bureau - Medical and Health Technology Project, No. Y20212003 (to ZGC); Precision Medicine Project of Wuxi Municipal Health Commission, No. J202002 (to ZGC)

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摘要：


文题释义：

面神经：第七对颅神经，属于外周神经系统，经茎乳孔出颅后分为5支，主要支配面部表情肌群，损伤后引起面瘫。
毛囊表皮神经嵴干细胞：毛囊隆突区来源的表皮神经嵴干细胞，具备多向分化潜能。起源于神经外胚层神经嵴，与外周神经具有同源性，是外周神经损伤修复极具潜力的种子细胞。

背景：既往研究报道了毛囊表皮神经嵴干细胞在周围神经修复中的巨大潜力，但其炎症调节作用在面神经修复研究中鲜有报道。
目的：探究毛囊表皮神经嵴干细胞能否调节面神经损伤后早期局部肿瘤坏死因子α和白细胞介素4表达水平，以促进面神经功能和形态恢复。
方法：提取4 d龄SD大鼠毛囊表皮神经嵴干细胞进行培养、鉴定。取54只成年雄性SD大鼠制备面神经干缺损自体静脉导管桥接模型，随机等分为生理盐水组、DMEM组和毛囊表皮神经嵴干细胞组。通过免疫蛋白印迹、免疫组化、免疫荧光观察术后4-14 d肿瘤坏死因子α和白细胞介素4表达水平。对术后 12 周大鼠进行面神经功能评分，并行苏木精-伊红染色观察面神经形态。

结果与结论：①毛囊表皮神经嵴干细胞中 Nestin 和 p75NTR 双阳性表达，细胞纯度 > 90%；②与其他两组相比，毛囊表皮神经嵴干细胞组的肿瘤坏死因子α表达在术后7 d减弱(P < 0.05)，而白细胞介素4表达在术后3，7，14 d均增强(P < 0.05)；③与其他两组相比，毛囊表皮神经嵴干细胞组的面神经功能改善(P < 0.05)；④面神经移植段均可见新生血管和再生轴突，与其他两组相比，毛囊表皮神经嵴干细胞组移植段和移植远段的神经纤维排列更有序、包绕的髓鞘更厚；⑤结果表明：毛囊表皮神经嵴干细胞对面神经损伤后修复早期局部肿瘤坏死因子α和白细胞介素4表达具有调节作用，可抑制局部炎症反应，并促进面神经功能和形态恢复。

https://orcid.org/0000-0003-4814-1550 (朱国臣) 

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: 面神经, 神经再生, 毛囊, 表皮神经嵴干细胞, 炎症因子, 肿瘤坏死因子α, 白细胞介素4

Abstract: BACKGROUND: Previous studies had shown that hair follicle epidermal neural crest stem cells have the great potential in facilitating peripheral nerve rehabilitation, but the role in regulating inflammation after facial nerve injury had rarely been reported.  
OBJECTIVE: To investigate whether epidermal neural crest stem cells can modify the expression of pro-inflammatory factor tumor necrosis factor α and anti-inflammatory factor interleukin-4 during facial nerve regeneration to promote the functional and morphological recovery of the facial nerve.
METHODS: Epidermal neural crest stem cells were extracted from one 4-day-old SD rat, cultured and identified. Fifty-four adult SD rats were used to construct the bridge model of facial nerve trunk defect with the autologous venous catheter. Models were randomly and equally divided into the normal saline group, Dulbecco's modified eagle medium group and epidermal neural crest stem cell group. Tumor necrosis factor α and interleukin-4 expression levels were detected at postoperative 4 to 14 days using western blotting, immunohistochemistry, and immunofluorescence. The facial nerve function of rats was scored 12 weeks after the operation and the morphology of the facial nerve was observed by hematoxylin-eosin staining.  
RESULTS AND CONCLUSION: (1) Double positive expression of Nestin and p75NTR could be observed in epidermal neural crest stem cells and the cell purity was over 90%. (2) Compared with the other two groups, tumor necrosis factor α expression in the epidermal neural crest stem cell group turned weaker 7 days after surgery (P < 0.05), and interleukin-4 expression in the epidermal neural crest stem cell group was enhanced on postoperative 3, 7 and 14 days, respectively (P < 0.05). (3) The recovery score of facial nerve function in the epidermal neural crest stem cell group was significantly better than that in the other two groups (P < 0.05). (4) There were abundant nutrient vessels and regenerative axons in the graft segment. Compared with the other two groups, the arrangement of nerve fibers was more orderly and the wrapped myelin sheath was thicker in the graft and distal segment in the epidermal neural crest stem cell group. (5) The data inferred that epidermal neural crest stem cells regulate the local expression of tumor necrosis factor α and interleukin-4 at the early stage of facial nerve injury, inhibit the local inflammatory reaction, and improve facial nerve function and morphology of the regenerated nerve.

Key words: facial nerve, nerve regeneration, hair follicle, epidermal neural crest stem cell, inflammatory factor, tumor necrosis factor α, interleukin-4

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