基于巨噬细胞M2型极化眶内电针干预下动眼神经功能重建机理的研究

Oculomotor nerve palsy(ONP), as a commonly seen but complicated cranial nerve neuropathy, is usually secondary to cerebral and metabolic disorders. The patient would be tortured physically and psychologically due to the manifestations include diplopia and ocular motility disorders. Nowadays, functional reconstruction of ONP has been highly-focused by researchers from international medical community. However, efficient rehabilitation method hasn’t been reported yet. Our team first adopt the intraocular electroacupuncture therapy into ONP rehabilitation and has treated 1032 patients since 2007. By statistical analysis, the results indicate that this therapy could remarkably relieving or even eliminating the symptoms and effectively improving the patient’s quality of life. In order to elucidate the actual mechanism of the therapy, we would use SD rat as the object to observe the macrophages M2 polarizing which play a notable role in nerve regeneration during intervening by intraocular electroacupuncture. We would apply western blot and RT-PCR technique to investigate the protein and mRNA expression level of immune reactive and neurotrophy factors, by which to elucidate the molecule mechanism of intraocular electroacupuncture interfereing the functional reconstruction of ONP. This project could not only provide scientific evidence for promoting the therapy into clinical practice, but also lay the technical foundation for upgrading the position of acupuncture in treating incurable diseases in medical world.

动眼神经麻痹（Oculomotor Nerve Palsy, ONP）是常见的脑神经疑难疾病，多继发于脑部病变和代谢性疾病，临床表现为复视、眼球运动障碍等，给患者造成巨大的身心痛苦。目前ONP功能修复已引起国际医学界的高度关注，然而，有效的西医康复手段尚未见报道。本课题组自2007年率先采用眶内电针方法治疗ONP患者1032例，初步分析显示：该方法可大幅改善甚至消除患者ONP症状，有效提高其生存质量。为从机理上阐明该方法的有效性，本项目拟采用雄性SD大鼠为实验动物，以神经再生过程中发挥重要作用的巨噬细胞M2型极化为研究目标，采用免疫荧光标记、蛋白印迹等方法进行检测，通过记录神经生长因子、免疫反应因子的蛋白及基因表达水平，从分子层面阐释眶内电针在ONP功能重建过程中的作用机制，为实现该方法在临床上的推广应用提供科学依据，同时，为提升针灸治疗疑难杂症在医学界中地位奠定必要的技术储备。

动眼神经损伤（Oculomotor Nerve Palsy，ONP）后所导致的双眼视觉偏差、眼球运动功能障碍，严重影响患者的心理健康和生活质量，给社会和家庭带来沉重负担。本课题组采用眶内电针疗法治疗ONP取得良好的临床疗效，本次研究主要目的为探寻眶内电针疗法起效的物质基础。本次研究首先探索大鼠动眼神经夹闭损伤模型的建造方法，提出了蝶腭孔入路这一新的术式成功建造了动眼神经损伤模型。模型创建成功后进行眶内电针疗效的验证，根据大鼠眼球运动距离与瞳孔直径等数据，确定了眶内电针对于大鼠动眼神经损伤模型的疗效。通过组织染色比较不同组别不同时间点神经轴突的形态与密度，结果数据说明眶内电针可以促进动眼神经损伤的修复。在此重要前提下本次研究使用免疫荧光，western blot，PCR以及流式细胞术等检测方法验证眶内电针干预后巨噬细胞及其相关蛋白的变化情况，从而验证眶内电针是否干预了受损组织内的巨噬细胞M2型的表型变化，以及M2型巨噬细胞下游神经生长因子等相关蛋白的结果，进一步验证了眶内电针起效的机制之一是通过巨噬细胞表型变化起作用。更深一步探索眶内电针是如何影响巨噬细胞表型变化，研究结果发现眶内电针可以干预组织内相关细胞因子的表达，从而干预微环境使其向有利于M2型细胞极化的组织修复环境转化。本次研究不仅体现中医针灸在神经损伤方面具有独特优势，还为中医经络腧穴作用理论研究积累相关基础，为针灸有效机制的研究增加必要的科学内涵。