从MMP9/IL-1β信号通路探讨蒙古族药那如-3对神经病理性疼痛“启动期”神经炎症的干预机制  

Mechanism of Mongolian drug Naru-3 in initiation of neuroinflammation of neuropathic pain from MMP9/IL-1βsignaling pathway

作　　者：周方婷 宗瑛 李元滨 曹人郦 侯吴琼 许丽婷 杨菲[2] 顾艳丽[4] 苏晓慧[2] 郭秋岩 李玮婕 熊慧[6] 王超[2] 林娜[1,2] ZHOU Fang-ting;ZONG Ying;LI Yuan-bin;CAO Ren-li;HOU Wu-qiong;XU Li-ting;YANG Fei;GU Yan-li;SU Xiao-hui;CUO Qiu-yan;LI Wei-jie;XIONG Hui;WANG Chao;LIN Na(School of Traditional Chinese Medicine,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Bejing 100700,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Bejing 100700,China;Inner Mongolia Medical University,Hohhot 010110,China;Artemisinin Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Pharmaceutical Sciences,South-Central Minzu University,Wuhan 430074,China)

机构地区：[1]广州中医药大学中药学院,广东广州510405 [2]中国中医科学院中药研究所,北京100700 [3]北京中医药大学东直门医院,北京100700 [4]内蒙古医科大学,内蒙古呼和浩特010110 [5]中国中医科学院青蒿素研究中心,北京100700 [6]中南民族大学药学院,湖北武汉430074

出　　处：《中国中药杂志》2023年第15期4173-4186,共14页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81673695,82174084,81873322);国家重点研发计划项目(2022YFC3502202);中国民族医药学会科研项目(2021Z1214-080302,2022M2122-070303,2022M2123-070304)。

摘　　要：神经病理性疼痛(neuropathic pain,NP)表型相似而病理过程的序贯性神经炎症机制各异,将神经炎症抑制在“启动期”具有重要意义并成为近年来NP治疗和药物研发的新方向。蒙古族药那如-3(Naru-3)临床治疗三叉神经痛、坐骨神经痛等NP短期即效,但其镇痛的药效学特点及机制尚不明确。该研究建立模拟临床周围神经损伤的脊神经结扎(spinal nerve ligation,SNL)模型并采用行为学检测、副作用评估、网络分析及实验验证等方法探讨Naru-3治疗NP的药效机制。药效学结果显示,Naru-3能提高SNL动物“启动期”基础痛敏阈值(机械痛敏、热辐射痛敏),缓解自发痛,而对正常动物基础痛敏阈值、生理及病理状态下动物的运动协调功能无明显影响。靶组织初筛结果表明Naru-3能抑制小鼠福尔马林实验第二时相伤害性反应并降低脊髓炎症因子表达。网络分析发现Naru-3治疗NP具有协同性且与MMP9、IL1B等核心靶标相关。实验进一步以背根神经节(dorsal root ganglia,DRG)、病变脊髓节段为研究对象,聚焦于诱导小胶质细胞神经炎症的核心靶标,采用免疫印迹、免疫荧光、激动剂、拮抗剂、行为学等技术方法发现Naru-3发挥NP镇痛作用的机制可能与其负向调节MMP9/IL-1β信号通路介导的“启动期”小胶质细胞p38/IL-1β炎症环路相关。相关研究丰富了Naru-3治疗NP的生物学内涵,并为其临床合理用药提供了参考。Neuropathic pain(NP)has similar phenotypes but different sequential neuroinflammatory mechanisms in the pathological process.It is of great significance to inhibit the initiation of neuroinflammation,which has become a new direction of NP treatment and drug development in recent years.Mongolian drug Naru-3 is clinically effective in the treatment of trigeminal neuralgia,sciatica,and other NPs in a short time,but its pharmacodynamic characteristics and mechanism of analgesia are still unclear.In this study,a spinal nerve ligation(SNL)model simulating clinical peripheral nerve injury was established and the efficacy and mechanism of Naru-3 in the treatment of NPs was discussed by means of behavioral detection,side effect evaluation,network analysis,and experimental verification.Pharmacodynamic results showed that Naru-3 increased the basic pain sensitivity threshold(mechanical hyperalgesia and thermal radiation hyperalgesia)in the initiation of SNL in animals and relieved spontaneous pain,however,there was no significant effect on the basic pain sensitivity threshold and motor coordination function of normal animals under physiological and pathological conditions.Meanwhile,the results of primary screening of target tissues showed that Naru-3 inhibited the second phase of injury-induced nociceptive response of formalin test in mice and reduced the expression of inflammatory factors in the spinal cord.Network analysis discovered that Naru-3 had synergy in the treatment of NP,and its mechanism was associated with core targets such as matrix metalloproteinase-9(MMP9)and interleukin-1β(IL-1β).The experiment further took the dorsal root ganglion(DRG)and the stage of patho-logical spinal cord as the research objects,focusing on the core targets of inducing microglial neuroinflammation.By means of Western blot,immunofluorescence,agonists,antagonists,behavior,etc.,the mechanism of Naru-3 in exerting NP analgesia may be related to the negative regulation of the MMP9/IL-1βsignaling pathway-mediated microglia p38/IL-1βinfl

关 键 词：蒙古族药那如-3 神经病理性疼痛神经炎症“启动期” 小胶质细胞活化 MMP9/IL-1β信号通路 机制 

分 类 号：R29[医药卫生—民族医学]