口服烟酰胺核糖抑制实验性自身免疫性脑脊髓炎的初步研究  

Preliminary study on inhibition of experimental autoimmune encephalo⁃myelitis by oral nicotinamide riboside

作　　者：宋国斌[1] 席国萍[1] 王青 章培军[1] 孟涛 刘春云[1] 尉杰忠[1,3] 肖保国 李艳花[1] 马存根[1,2] SONG Guobin;XI Guoping;WANG Qing;ZHANG Peijun;MENG Tao;LIU Cunyun;YU Jiezhong;XIAO Baoguo;LI Yanhua;MA Cungen(Institute of Brain Science,Shanxi Datong University,Datong 037009,China;The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine,Shanxi University of Chinese Medicine,Jinzhong 030619,China;The Fourth People's Hospital of Datong,Datong 037000,China;Institute of Neurology,Huashan Hospital,Fudan University,Shanghai 200025,China)

机构地区：[1]山西大同大学脑科学研究所,山西大同037009 [2]山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室,山西晋中030619 [3]大同市第四人民医院,山西大同037000 [4]复旦大学华山医院神经病学研究所,上海200025

出　　处：《中国病理生理杂志》2023年第4期608-615,共8页Chinese Journal of Pathophysiology

基　　金：山西省基础研究计划项目(No.202203021211327);国家自然科学基金资助项目(No.81903596);基于炎性反应的重大疾病创新药物山西省重点实验室项目(No.202105D121011);药用资源与天然药物化学教育部重点实验室开放课题(No.2019004);山西省卫健委医学科技领军团队(No.2020TD05);国家中医药管理局多发性硬化益气活血重点研究室经费;山西中医药大学学科建设经费。

摘　　要：目的:探讨口服烟酰胺核糖(nicotinamide riboside,NR)对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的治疗效果和作用机制。方法:采用髓鞘少突胶质细胞糖蛋白35-55(myelin oligodendrocyte glycoprotein 35-55,MOG_(35-55))免疫C57BL/6雌性小鼠制备EAE模型,随机分为EAE组和NR组。从免疫后第3天开始,EAE组小鼠灌胃给予生理盐水,NR组小鼠灌胃给予NR,每天1次至免疫后第27天。免疫当天至免疫后第28天,观察并记录各组小鼠临床评分和体重。免疫后第28天处死小鼠,制备脾细胞悬液和脊髓组织冰冻切片。HE染色和髓鞘染色分别检测脊髓炎性细胞浸润和髓鞘脱失,Western blot和免疫荧光染色分别检测脊髓组织中ROCK-II、TLR4、p-NF-κB、iNOS表达及相应的阳性细胞数量,ELISA检测脾细胞培养液中TNF-α、IL1β和IL-6的含量。结果:口服NR可推迟EAE发病时间,减轻EAE临床症状,缓解小鼠体重丢失,减少外周炎性细胞浸润脊髓,减轻髓鞘脱失,降低脊髓组织中ROCK-II、TLR4、p-NF-κB及iNOS炎性相关蛋白表达,抑制脾脏炎性因子分泌。结论:口服NR对EAE具有显著的治疗效果,其治疗机制可能与NR抑制EAE的炎症反应有关。AIM:To explore the therapeutic effect and mechanism of oral nicotinamide riboside(NR)on experimental autoimmune encephalomyelitis(EAE).METHODS:Female C57BL/6 mice immunized with myelin oligodendrocyte glycoprotein 35-55(MOG_(35-55))were randomly divided into EAE group and NR group.From the 3rd day to the 27th day after immunization,the mice in EAE group were given normal saline and those in NR group were given NR once a day by intragastric administration.Clinical score and body weight of mice in each group were recorded from the day of immuni·608·zation to the 28th day after immunization.On the 28th day after immunization,mice in each group were sacrificed and splenic cell suspension and frozen sections of spinal cord were prepared.Inflammatory cell infiltration and myelin demyelination were detected separately by HE staining and LFB staining.The expression of ROCK-II,TLR4,p-NF-κB,iNOS and the number of corresponding positive cells in spinal cord were respectively detected by Western blot and immunofluorescence staining.The contents of TNF-α,IL-1βand IL-6 in splenic cell culture medium were determined by ELISA.RE⁃SULTS:Oral administration of NR delayed the onset of EAE,attenuated the clinical symptoms of EAE,relieved weight loss of mice,decreased the infiltration of peripheral inflammatory cells into spinal cord,reduced myelinoclasis,lessened the expression levels of inflammation-related proteins such as ROCK-II,TLR4,p-NF-κB and iNOS,and inhibited the secretion of inflammatory factors in spleen.CONCLUSION:Oral NR has a significant therapeutic effect on EAE,and its therapeutic mechanism may be related to the inhibition of the inflammatory response of EAE.

关 键 词：烟酰胺核糖 实验性自身免疫性脑脊髓炎 RHO/ROCK信号通路 TLR4/NF-κB信号通路 

分 类 号：R744.51[医药卫生—神经病学与精神病学] R364.5[医药卫生—临床医学]