中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (8): 1253-1259.doi: 10.12307/2022.232

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

电针干预面神经损伤模型兔面神经核团中神经生长因子及其受体的表达

肖  阳,龚立琼,费  静,李雷激   

  1. 西南医科大学附属医院耳鼻咽喉头颈外科,四川省泸州市   646000
  • 收稿日期:2020-12-01 修回日期:2020-12-16 接受日期:2021-01-16 出版日期:2022-03-18 发布日期:2021-11-02
  • 通讯作者: 李雷激,硕士,主任医师,西南医科大学附属医院耳鼻咽喉头颈外科,四川省泸州市 646000
  • 作者简介:肖阳,男,1993年生,四川省广安市人,汉族,西南医科大学在读硕士,主要从事面神经损伤的基础与临床研究。

Effect of electroacupuncture on nerve growth factor and its receptor expression in facial nerve nucleus after facial nerve injury in rabbits

Xiao Yang, Gong Liqiong, Fei Jing, Li Leiji   

  1. Department of Otorhinolaryngology Head and Neck Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2020-12-01 Revised:2020-12-16 Accepted:2021-01-16 Online:2022-03-18 Published:2021-11-02
  • Contact: Li Leiji, Master, Chief physician, Department of Otorhinolaryngology Head and Neck Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Xiao Yang, Master candidate, Department of Otorhinolaryngology Head and Neck Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China

摘要:

文题释义:
神经生长因子受体:是一类与神经营养因子特异性结合的生长因子受体,包括低亲和力神经生长因子受体(p75等)和属于受体酪氨酸激酶的TRK受体家族。p75是一种低亲和力神经生长因子受体,可以与所有神经营养因子结合,是肿瘤坏死因子受体超家族的一员。
酪氨酸激酶(TrkA):是神经营养因子的功能性受体,当其与神经营养因子结合后,可激活酪氨酸激酶信号传递系统,从而启动细胞活性,产生生物效应。
背景:研究发现周围性面神经损伤能引起神经生长因子表达变化,而电针穴位治疗对神经生长因子的影响机制尚不清楚。
目的:通过建立兔周围性面神经损伤模型,予以电针刺激后,观察其对兔面瘫症状和面神经核团形态学的影响以及对神经生长因子及其受体表达的影响,探讨电针在周围性面神经损伤治疗中的作用机制。
方法:将42只新西兰大白兔随机分为正常组(n=6)、模型组(n=18)、电针组(n=18)。正常组不做处理,其他2组兔建立右侧面神经上颊支压榨损伤模型。造模成功后,电针组每日取“合谷”穴埋针,“颊车”“地仓”“四白”“阳白”“翳风”“颧髎”6个穴位予以疏密波刺激;模型组不作治疗。电针治疗28 d采用苏木精-伊红染色观察面神经核团的形态学,免疫组织化学染色检测面神经核团中神经生长因子及其受体 TrkA 和 p75NTR 的表达部位及强度,并通过荧光定量 PCR 法检测面神经核团中神经生长因子及其受体mRNA 的表达。实验经西南医科大学动物伦理学委员会批准,批准号为20170120001。 
结果与结论:①苏木精-伊红染色可见:电针组神经元术后形态恢复较模型组快且完全;②免疫组织化学染色显示:术后各时间点电针组神经生长因子的平均吸光度均高于模型组(P < 0.05),电针组TrkA、p75NTR的平均吸光度在术后 7,14,28 d均高于模型组(P < 0.05);③荧光定量 PCR 检测结果显示:术后 14,28 天,电针组中神经生长因子和p75NTR mRNA 相对表达量高于模型组;术后 7,14,28天,电针组TrkA mRNA 的相对表达量高于模型组(P < 0.05);④结果提示:电针可以提高面神经核团中神经生长因子、TrkA、p75NTR的表达,并可能促进神经生长因子与 TrkA和 p75NTR的分别结合,这可能是电针在促进面神经损伤后修复的机制之一。

https://orcid.org/0000-0002-6598-8019 (肖阳) 

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程

关键词: 电针, 穴位, 周围性面瘫, 面神经核团, 神经生长因子, p75 神经生长因子受体, 神经生长因子络氨酸激酶受体 A

Abstract: BACKGROUND: Preliminary studies have found that peripheral facial nerve injury can cause changes in the expression of nerve growth factor, but the mechanism of electroacupuncture in the expression of nerve growth factor is unclear.
OBJECTIVE: Based on a model of peripheral facial nerve injury in rabbits, to observe the effects of electroacupuncture on facial paralysis symptoms, morphology of facial nerve nuclei and expression of nerve growth factor and its receptor and to explore the mechanism of electroacupuncture in the treatment of peripheral facial nerve injury. 
METHODS: Forty-two New Zealand white rabbits (n=66) were randomized into normal group (n=6), model group (n=30) and electroacupuncture group (n=30). A crushing injury model of superior buccal branch of the right facial nerve was established in the latter two groups. There was no deal with normal group. After successful modeling, a needle was imbedded at Hegu in the electroacpuncture group, followed by electric stimulations with dilatational wave at Jiache, Dicang, Sibai, Yangbai, Yifeng, and Quanliao. No treatment was done in the model group. At 28 days of electroacupuncture, morphological changes of facial nerve nuclei were observed using hematoxylin-eosin staining and the expression of nerve growth factor, TrkA and p75NTR in facial nerve nuclei was detected using immunohistochemical staining. The expression of nerve growth factor, TrkA and p75NTR mRNA was detected using fluorescent quantitative PCR. The study protocol was approved by the Animal Ethics Committee of Southwest Medical University, with an approval No. 20170120001.
RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that the morphology of neurons recovered faster and more completely in the electroacupuncture group than the model group after surgery. The results of immunohistochemical staining showed that the average absorbance value of nerve growth factor in the electroacupuncture group was higher than that in the model group at every time point (P < 0.05), and the average absorbance value of TrkA and p75NTR in the electroacupuncture group was higher than that in the model group on the 7th, 14th, and 28th days after surgery (P < 0.05). Fluorescent quantitative PCR results revealed that the relative expression of nerve growth factor and p75NTR mRNA in the electroacupuncture group was higher than that in the model group on the 14th and 28th days after surgery (P < 0.05). The TrkA mRNA expression in the electroacupuncure group was higher than that in the model group on the 7th, 14th, and 28th days after surgery (P < 0.05). To conclude, electroacupuncture can improve the expression of nerve growth factor, TrkA, and p75NTR in facial nerve nuclei, and may promote the binding between nerve growth factor with TrkA and p75NTR, which may be the mechanism by which electroacupuncture promotes the repair of facial nerve injury.


Key words: electroacupuncture, acupoint, peripheral facial paralysis, facial nerve nucleus, nerve growth factor, p75 nerve growth factor receptor, nerve growth factor tyrosine kinase receptor A

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