水通道蛋白1在BELL's面瘫神经水肿损伤中的作用和调节机制

Bell's palsy often leads to sustained or permanent facial deformity and psychological barriers. Research had showed that, the facial nerve edema and hoop pressure from the facial canal is the direct etiological factor, but the report related to the pathogenesis had not been seen. In recent years, the aquaporins (AQPs) had been found closely related with edema diseases. Our research group had found that AQP1 distributed in facial nerve abundantly, and expression increased after the nerve injury. Is AQP1 related to the edema pathogenesis of Bell's palsy, or whether the regulation of AQP1 expression can alleviate nerve edema? Firstly, our reserch group will establish the HSV-1-induced mouse facial paralysis model, detect the distribution and change of AQP1 expression by the mothods as Immunohistochemistry, RT-PCR and so on. Then, Combined with cell culture and identification, explicit the positioning of AQP1 completely at organism and cellular levels. Secondly, at cellular level, by using the regulation function of acetazolamide to AQP1, adopting gene-silencing technology, and testing the value of the osmotic water permeability (Pf), we will explore the relationship of AQP1 and nerve edema, and further clear the regulation mechanism of AQP1. At last, we will apply acetazolamide to the facial paralysis model, measure the facial nerve/facial canal (FN/FC) ratio and the canal pressure, and count the numbers of the apoptosis facial motorneuron. Then, compare the results from the different experimental groups to evaluate the influence of acetazolamide to the facial nerve edema and the prognosis by regulating the AQP1 expression.This study explore the relationship and mechanism of AQPs and the nerve edema injury in Bell's palsy for the first time, and will provide a new selection drug for clinical treatment.

BELL's面瘫常导致持续或永久的面部畸形和心理障碍。研究表明，面神经水肿和面神经管箍压是其直接病因，但尚未见相关发病机制的报道。近年发现，水通道蛋白（AQPs）与水肿性疾病密切相关。本课题组前期研究发现AQP1分布于面神经周围，且在神经损伤后表达增强。AQP1与BELL's面瘫神经水肿发生是否相关，能否通过调节AQP1的表达来减轻神经水肿？本课题拟构建HSV-1诱导小鼠面瘫模型，通过免疫组化和RT-PCR等检测AQP1表达，结合细胞培养鉴定，完成AQP1组织细胞学定位；通过乙酰唑胺药物调控和基因沉默，结合透水性(Pf)测定，明确AQP1与神经水肿的因果关系及调节机制；将乙酰唑胺应用于面瘫模型，测定FN/FC值和管内压，对比面神经核团凋亡情况，评估其通过调节AQP1表达对面神经水肿及预后的影响，本课题首次探讨AQPs与BELL's面瘫面神经水肿损伤的关系及机制，拟为临床提供理想的治疗方法。

BELL’ s面瘫常导致持续或永久的面部畸形和心理障碍。研究表明，面神经水肿和面神经管箍压是其直接病因，但尚未见相关发病机制的报道。近年发现，水通道蛋白（ AQPs）与水肿性疾病密切相关。本课题成功建立稳定、可靠的特发性面神经麻痹动物模型，采用多重免疫荧光双标和细胞培养鉴定等技术方法，首次发现AQP1分布于面神经颞骨部的神经组织和神经滋养血管中，定位于雪旺氏细胞、血管内皮细胞及神经外膜，并且在建立的特发性面神经麻痹动物模型中发现AQP1mRNA和蛋白表达水平呈急起缓降的趋势，并与HSV-1诱导急性可复性面神经麻痹后神经水肿的变化趋势相吻合；通过应用RNA干扰和过表达技术分别下调和上调雪旺细胞AQP1表达，发现AQP1是控制雪旺细胞快速水转运的主要因素，是参与雪旺细胞可塑性的一种蛋白；通过制作雪旺细胞CoCl2缺氧模型，模拟体内面神经损伤，研究在缺氧病理状态下AQP1的表达变化及其对雪旺细胞形态和水转运的影响，发现AQP1表达变化是缺血、缺氧诱导面神经水肿的主要因素，HIF-1α参与缺氧诱导AQP1表达增高，为临床预防或减轻面神经水肿后的继发损伤提供理论依据。并且在细胞和动物层面发现乙酰唑胺可抑制AQP1的表达和功能，降低面神经水肿峰值的程度，提示在面神经损伤水肿过程中，早期应用乙酰唑胺可减轻面神经的水肿。本课题首次探讨 AQPs 与 BELL's 面瘫面神经水肿损伤的关系及机制，为临床治疗提供理论基础。