缓释CXCL12对受损面神经雪旺氏细胞迁移及神经修复的影响与机制研究

Facial spasm is a common cranial nerve disease, seriously affecting the quality of life. Vascular compression facial nerve causes facial nerve demyelinating changes is the main basis, and in microvascular decompression surgery, it is the main reason of facial nerve demyelinating lesions can't complete repair for invalid surgery and recurrence of symptoms postoperatively. At the present stage it is lack of effective ways to promote restoration of the facial nerve demyelination. This experiment is to be established in the rabbit facial nerve demyelinating animal models as well as the synthesis of RADA16-I hydrogel fixed with chemotactic factor CXCL12, on the basis of the migration experiment (Transwell), Stripe (Stripe), flow cytometry, quantitative detection of apoptosis (TUNEL) technique, neural function electrophysiological evaluation, neuroanatomy and various experimental techniques to observe the influence of in vitro and in vivo slow release of CXCL12 on schwann's cell migration and the facial nerve demyelination repair. On the basis of the analysis of the local microenvironment with cytokine chip and the method of intervention CXCR4/7 pathways to study the molecular mechanism of the CXCL12 promote restoration of the facial nerve injury. The completion of this study is expected to provide theoretical support for developing clinical new techniques on facial nerve repair and functional recovery.

面肌痉挛是一种常见的颅神经疾病，严重影响生活质量。血管压迫面神经导致面神经脱髓鞘改变是主要的发病基础，微血管减压术后面神经脱髓鞘损伤不能完全修复是术后无效和症状复发的主要原因，目前临床上尚缺乏促进面神经脱髓鞘修复的有效方法。本实验将在建立实验兔面神经脱髓鞘动物模型以及合成转载趋化因子CXCL12的纳米水凝胶RADA16-I的基础上，采用迁移实验（Transwell）、条带实验（Stripe）、流式细胞技术定量检测细胞凋亡(TUNEL)、神经功能电生理学评估、神经组织学等多种实验技术观察体外和体内环境下缓慢释放CXCL12对雪旺氏细胞迁移和面神经脱髓鞘修复方面的作用，并采用细胞因子芯片解析受损面神经局部微环境以及干预CXCR4/7通路的方法研究CXCL12促进面神经损伤修复的分子机制。本实验的完成有望为开发促进面神经损伤修复及功能恢复的临床新技术提供理论支持。

面肌痉挛是一种常见的颅神经疾病，严重影响生活质量。血管压迫面神经导致面神经脱髓鞘改变是主要的发病基础，微血管减压术后面神经脱髓鞘损伤不能完全修复是术后无效和症状复发的主要原因，目前临床上尚缺乏促进面神经脱髓鞘修复的有效方法。雪旺氏细胞在面神经损伤后通过迁移到损伤部位，并在修复神经损伤的过程发挥重要作用。CXCL12作为一种趋化相关因子和细胞迁移密切相关。但CXCL12在面神经损伤后的作用尚不清楚。本实验建立大鼠面神经损伤模型模拟面神经脱髓鞘损伤，通过迁移实验、电镜技术、流式细胞技术、细胞凋亡荧光染色、神经功能电生理学评估、神经组织学等多种实验技术观察体外和体内环境下CXCL12对雪旺氏细胞迁移和面神经脱髓鞘修复方面的作用。实验结果表明CXCL12在面神经损伤早期表达上调，两周后表达降低。进一步研究发现CXCL12对雪旺细胞增殖、凋亡和细胞周期无影响，但显著促进雪旺细胞迁移。应用CXCL12处理后可降低雪旺氏细胞PI3K、AKT和mTOR磷酸化，但自噬标记物LC3II/I增加。通过自噬抑制剂3-MA预处理抑制自噬，以及IGF-1激活PI3K通路，发现CXCL12诱导的雪旺细胞迁移显著减弱，而PI3K通路抑制剂LY294002可以增强这种作用。动物实验也证实CXCL12能改善面神经功能和髓鞘再生。综上，研究结果证明CXCL12通过PI3K-AKT-mTOR信号通路促进自噬，进而促进雪旺细胞迁移，从而改善面神经髓鞘再生和面神经功能。因此本实验有望为开发促进面神经损伤修复及功能恢复的临床新技术提供理论支持，CXCL12有可能成为修复面神经损伤的关键分子。