管灸疗法促面神经损伤修复的NT-Trk信号转导机制研究

Guan moxibustion is an unique moxibustion method with long history and distinctive characteristics of Traditional Chinese Medicine .It is the first batch of TCM clinical extension technology by the State Administration of traditional Chinese medicine.It has has prominent treatment effect with facial nerve injury,and has the characteristics of simple operation, no injury and painless , patients vulnerable to accept and high compliance. Research group has Guan moxibustion patent and has found that Guan moxibustion can enhance expression level of nerve nutrition ( NT ) and Trk receptors in the nucleus of facial nerve, facial nerve, facial muscle. NT is considered to play an important role in the repair process of damaged facial nerve. NT/Trk signal transduction pathways is a main signal pathways which NT play effections of nerve repairing and regenerating, the subject closely surround the working hypothesis which Guan moxibustion can promote reparation of the damaged facial nerve is due to increasing expression level of NT and Trk receptors , activating NT/Trk signal transduction pathway ,use integratedly electrophysiology, transmission electron microscopy, immunohistochemistry, in situ hybridization and RT-PCR technology to discover the Guan moxibustion's regulating mechanism on NT/Trk signal transduction pathway. The results will clarify modern scientific principle of Guan moxibustion in the therapy of facial nerve injury, and provide for its clinical application more fully scientific basis, enrich and develop moxibustion theory.

管灸是具有悠久历史和鲜明中医特色的独特灸法，为国家中医药管理局第一批中医临床推广技术，治疗面神经损伤疗效突出，操作简便，无创无痛，患者依从性高。课题组拥有改进的管灸器专利，并且研究发现管灸能增强面神经核、面神经、面部表情肌中神经营养素（NT）及其Trk受体的表达。目前认为NT在受损面神经的修复过程中起着不可或缺的重要作用，而NT/Trk信号转导通路是NT发挥神经修复再生效应的最主要信号转导通路，本课题紧密围绕"管灸促面神经损伤修复主要是通过增强NT及其Trk受体表达进而激活NT/Trk信号转导通路而实现的"工作假说，综合运用电生理、透射电镜、免疫组化、原位杂交及RT-PCR多种方法技术，探讨管灸对NT/Trk信号转导通路的调控作用。研究结果将阐明管灸疗法治疗面神经损伤的现代科学原理，为扩展其临床运用提供更为充分的科学依据，丰富发展灸法理论。

以面神经压榨法制备兔面神经损伤模型，运用光镜（HE染色和特殊染色）、电镜、电生理、免疫组化、蛋白印迹、原位杂交以及荧光定量PCR、生化检测等技术方法，围绕管灸疗法对面神经损伤治疗作用的NT-Trk信号转导机制这一研究目标，主要观察了临床症状改善情况，损伤面神经的组织病理学改变，电生理改变，与神经修复密切相关的NT/Trk信号通路中各关键环节信号转导分子的变化等。研究结果表明管灸疗法能够改善兔面神经的损伤家兔面瘫的临床症状，减轻组织病理学损伤，促进家兔面神经损伤的修复和再生。其机理与提高相应神经生长因子浓度及其Trks受体表达，并进一步升高神经营养因子胞内Ras/MEK/MAPK、PI3K/Akt、PLC信号转导通路的信号转导，增加与抗凋亡和促进细胞增殖和生长相关基因的表达而发挥促损伤面神经修复作用有关。研究结果基本阐明了管灸治疗面神经损伤的NT-Trk信号转导机制。同时单纯热疗对临床症状仅有部分改善作用，仅对部分与神经修复再生的相关因子有一定的影响，因此可以认为管灸疗法无论在临床疗效、作用靶点数量和对相应靶点的调节幅度上，均明显优于热疗，其作用不仅仅是依靠单纯热效应而起作用的，这进一步证明了管灸疗法的独特性。