SHARPIN介导的NF-κB信号通路的激活在单纯疱疹病毒感染性面瘫中的作用机制

It has been documented that infection of Herpes simplex virus type I (HSV-I) contributes to the initiation of Bell's palsy, to date, however, the exact mechanisms responsible for this disorder have not been fully elucidated. Previously, we found that the nuclear factor-κB (NF-κB) was activated in the facial nucleus of brainstem of the paralyzed mice elicited by HSV-I, but the mechanisms about how NF-κB was activated and whether its activation regulates the expressions of certain key target genes in this progress of facial paralysis are still unclear. On the basis of our previous researches, the present project is designed, utilizing HSV-I-induced Bell's palsy of mice and, simultaneously, the primary cultures of facial nucleus cells from brainstem of these infected mice, as the experimental models, to investigate the effects of alteration in activities of several inflammatory factors initiated by the activation of NF-κB pathway, on mouse facial palsy. In particular, this work will focus on exploring the mechanisms underpinning the action of SHANK-associated RH domain-interacting protein (SHARPIN) on linear ubiquitination of the NF-κB essential modulator (NEMO) and, consequently, leading to the activation of NF-κB. Our new research will lends insight into the etiology and mechanisms responsible for the Bell's palsy induced by HSV-I, which may, in turn, offer novel effective therapeutic targets to Bell's palsy.

I型单纯疱疹病毒(HSV-I)感染可导致贝尔面瘫，但其致病机制不明。我们前期研究发现，HSV-I感染致贝尔面瘫小鼠的面神经核团中核转录因子κB (NF-κB)被激活，但是目前关于NF-κB信号通路在HSV- I感染性面瘫中的激活机制及其激活后调控靶炎症因子的表达从而参与面神经炎症反应的作用及作用机制尚未见报道。本项目在前期工作的基础上，拟分别通过HSV-I感染致贝尔面瘫的小鼠动物模型及体外原代培养HSV-I感染性面瘫小鼠的脑干面神经核神经元细胞模型，研究NF-κB信号通路的激活导致的多种炎症因子的活性改变与HSV-I感染致贝尔面瘫的关系，并进一步深入探讨SHANK-相关RH结构域反应蛋白（SHARPIN）导致NF-κB必需调节蛋白（NEMO）泛素化后激活NF-κB信号通路的机制, 从而为HSV-I感染导致贝尔面瘫的作用机制研究提供新的理论依据, 进一步为贝尔面瘫的治疗提供新的作用靶点。

I型单纯疱疹病毒(HSV-I)感染可导致贝尔面瘫，但其致病机制不明。本项目在前期研究基础上，着重探讨了HSV-I感染致贝尔面瘫小鼠的面神经核团中核转录因子κB (NF-κB)信号通路在HSV- I感染性面瘫中的激活情况，及其激活后调控靶炎症因子的表达从而参与面神经炎症反应的作用和作用机制，阐述了SHANK-相关RH结构域反应蛋白（SHARPIN）在HSV- I感染性面瘫小鼠面神经核团中的表达改变情况，以及糖皮质激素通过调控上述因子表达从而治疗HSV-I感染导致的贝尔面瘫的作用及作用机制。本项目主要通过HSV-I感染致贝尔面瘫的小鼠动物模型，采用RT-PCR、western blot、EMSA等方法，发现HSV-I感染介导的面瘫小鼠脑干面神经核团中NF-κB信号通路可被激活并使其下游靶基因的表达出现时间依赖性的改变；揭示了在面瘫小鼠面神经核团中NF-κB可对TNF-α及COX-2的表达进行调控；阐明了在HSV-I诱导的面瘫小鼠脑干面神经核团中SHARPIN、单核细胞趋化因子-1(MCP-1)、细胞间粘附因子-1(ICAM-1)等因子可出现随时间依赖性的表达变化。并且通过进一步的实验证实，糖皮质激素MPSS可以改善HSV-I感染介导的面瘫小鼠的症状，其机制可能是通过抑制NF-κB、SHARPIN、MCP-1、ICAM-1在面瘫小鼠脑干面神经核团中的表达，以及由于抑制NF-κB表达而进一步调控了TNF-α和COX-2的表达实现的。本项目的研究为HSV-I感染导致贝尔面瘫的作用机制研究提供了新的理论依据, 进一步为贝尔面瘫的治疗提供了新的作用靶点。