Tmem100调控TRPV1-TLR4神经免疫复合体在慢性盘源性疼痛中的作用机制研究

椎间盘退变引起的腰背痛是骨科最常见的疼痛之一，73%的人一生中会出现严重的盘源性腰背痛，治疗效果不确切，极易迁延不愈。我们前期转染TAD多肽表达载体至大鼠背根神经节（DRG），可特异性干扰TRPV1 C端结构域蛋白结合位点功能，同时发现与肠道神经系统发育有关的Tmem100在DRG神经元中有表达，且与TRPV1及TLR4均存在共同表达。因此提出假设Tmem100作为一种重要的疼痛调节蛋白，与TRPV1及TLR4形成神经免疫复合体，在慢性盘源性疼痛中起着重要的作用。从细胞及动物模型两方面入手，借助免疫共沉淀、电生理等技术展开实验，明确Tmem100与TRPV1、TLR4特异性结合位点，阐明Tmem100与TRPV1-TLR4形成的神经免疫复合体在慢性盘源性疼痛中的表达及作用，并通过TAD多肽竞争抑制神经免疫复合体形成，探明其缓解慢性盘源性疼痛并调控免疫应答的机制，为盘源性疼痛治疗提供新思路。

Low back pain caused by disc degeneration was one of the most common problems in Department of Orthopaedics.73% of people had severe discogenic low back pain in his or her life with uncertain therapeutic efficacy and delayed recovery. In the pre-experiment, we had constructed an expression vector with functional TAD peptide, and then it was transfected to dorsal root ganglion(DRG) of the rats with the function of interference of C terminus of TRPV1.Tmeme100, which was related to the development of intestinal nervous system, was found to be expressed in DRG neuron, and was co-localized with TRPV1 and TLR4.Therefore, we proposed a hypothesis that Tmem100 as an important pain-regulating protein, formed a neuroimmune complex with TRPV1 and TLR4, and contributed to chronic discogenic pain. We aimed to start the experiment around cell and animal models, using many methods and techniques such as immunoprecipitation, electrophysiology, etc. We will clarify the specific binding sites of Tmem100 with TRPV1 and TLR4, and the expression and function of Tmem100 and TRPV1-TLR4 neuroimmune complex in chronic discogenic pain, and then demonstrate the mechanism of competitive inhibition of the neuroimmune complex for pain relief and regulation of the immune response after TAD transfection. We firmly believed that it will be valuable to provide a new way for treatment of chronic discogenic pain in the future.