针刺调控趋化因子FKN/CX3CR1通路抑制视网膜小胶质细胞活性效应研究

The loss of visual function caused by photoreceptor apoptosis is the final outcom in most retinal diseases is inreversible.After photoreceptor cells damaged,retinal microglia are activated,to sawllow, and damage by the chemokines FKN/CX3CR1 pathway accelerate their apoptosis.The topic is based on preliminary results of the study of acupuncture can inhibit the apoptosis of photoreceptor cells ,promote apoptosis of photoreceptor cells and visual function recovery.Intends to use the principle of acupuncture treatment,take chemokines FKN/CX3CR1 as the breakthrough point,use the ERG,Western blot,RT-PCR,immunoflourescence staining and other techniques,to explore the effect of acupuncture on the activity of microglia and the regulation of chemokines FKN/CX3CR1 and the expression of its downstream factors P42,P38,P-SAPK and so on,to clarify the biological effects of acupuncture on the activity of microglia,and its regualation approach and mechanism,to clarify the scientific connotation of acupuncture blocks apoptosis of retinal photoreceptor cells.Obtain empirical evidence of microglia as target to promote the survival of photoreceptor cells.Finally,it provides a new research idea for the prevention and treatment of photoreceptor apoptosis.

感光细胞凋亡是视网膜神经退行性疾病关键致病基础，视网膜小胶质细胞活化是促感光细胞凋亡的重要机制，前期研究证实针刺能抑制感光细胞凋亡、促进视功能恢复，但缺乏针刺对小胶质细胞活性效应的研究，本课题拟以前期证实针刺能抑制感光细胞凋亡为基础，瞄准诱导视网膜小胶质细胞活化、吞噬损伤感光细胞关键通路趋化因子FKN/CX3CR1信号系统，采用ERG、Westernblot、RT-PCR、免疫荧光染色等技术，探索针刺抑制小胶质细胞活性进而阻断感光细胞凋亡进程的确切效应，明确针刺调控趋化因子FKN/CX3CR1信号通路及其下游因子P38、P42、P-SAPK抑制视网膜小胶质细胞活化的关键机制，阐明针刺阻断视网膜感光细胞凋亡进程的科学内涵，获得以视网膜小胶质细胞为作用靶点保护感光细胞的针刺原理和实证依据，建立视网膜感光细胞凋亡的针灸干预评价方法，为针灸防治视网膜退行性疾病研究奠定实验基础，拓展新的研究领域。

感光细胞凋亡是视网膜退行性疾病(Retinal Degenerative,RD)关键致病因素，视网膜小胶质细胞活化在促感光细胞凋亡发生发展中发挥重要作用。针刺能抑制感光细胞凋亡、促进视功能恢复，但缺乏针刺对小胶质细胞活性效应的研究。本课题以MNU诱导的感光细胞凋亡大鼠为研究对象，结合体外实验，综合应用病理形态学、TUNLE、分子生物学、免疫荧光、Annexin V/PI 流式细胞术等研究方法，观察针刺对FKN/CX3CR1及JAK2/STAT3、NF-κB通路及视网膜小胶质细胞活性的影响，以阐明针刺治疗RD类疾病的科学内涵。主要研究结果表明：（1）针刺可以改善MNU诱导感光细胞凋亡大鼠的视网膜组织损伤，减缓或阻断感光细胞凋亡；（2）针刺能够抑制MNU大鼠视网膜小胶质细胞活化标志物IBa-1的表达及迁移，减轻炎症和感光细胞凋亡；（3）针刺可下调视网膜TNF-α、IL-1β、IL-6炎症因子释放，缓解MNU大鼠视网膜炎症反应；（4）针刺可下调大鼠视网膜FKN/CX3CR1信号通路关键分子表达，调控神经元-小胶质细胞相互作用，保护感光细胞凋亡；（5）针刺可通过JAK2/STAT3及NF-κB信号通路改善MNU大鼠视网膜损伤，抑制小胶质细胞活性，发挥减缓或阻断感光细胞凋亡进程的重要角色。系列研究揭示了针刺抑制小胶质细胞活性效应及其调控机制，为针刺以小胶质细胞为作用靶点，防治感光细胞凋亡提供了研究思路和理论依据。