灵孢多糖通过抑制TLR4/NF-κB通路改善EAE小鼠的髓鞘脱失  

GLPS improves EAE demyelination through inhibition of TLR4/NF-κB pathway

作　　者：李彦青 王青 杨智超 宋丽娟 王翰斌 杨立志 刘建春 肖保国[3] 马存根 LI Yan-qing;WANG Qing;YANG Zhi-chao;SONG Li-juan;WANG Han-bin;YANG Li-zhi;LIU Jian-chun;XIAO Bao-guo;MA Cun-gen(The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine,Research Center of Neurobiology,Shanxi University of Chinese Medicine,Jinzhong Shanxi 030619,China;Shanxi Guorun Pharmaceutical Co.,Ltd,Datong,Shanxi 038100,China;State Key Laboratory of Neurobiology,Dept of Neurology,Huashan Hospital,Fudan University,Shanghai 200025,China)

机构地区：[1]山西中医药大学神经生物学研究中心,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 [2]山西国润制药有限公司,山西大同038100 [3]复旦大学附属华山医院神经病研究所,上海200025

出　　处：《中国药理学通报》2023年第10期1914-1920,共7页Chinese Pharmacological Bulletin

基　　金：博士来晋科研启动项目(No 2021BKS01);山西省横向课题(No 2022HX-GR-01);山西中医药大学校级项目(No 2021PY-JC-15,2022BK15);山西省卫健委科技领军团队(No 2020TD05)。

摘　　要：目的探究灵孢多糖(ganoderma lucidum polysaccharides,GLPS)对多发性硬化(multiple sclerosis,MS)动物模型实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠的保护作用及机制。方法将C57BL/6小鼠30只随机平均分为3组:正常对照组、EAE模型组、GLPS组(5 mg·kg^(-1))。采用结核分枝杆菌和MOG35-55建立小鼠EAE模型,观察GLPS对EAE小鼠体质量及神经功能障碍评分的影响;采用固蓝染色和HE染色观察小鼠脊髓冰冻切片髓鞘组织病理情况、免疫荧光观察MBP表达变化;利用流式细胞术检测小鼠脾脏、淋巴结中IL-17、Foxp3含量;通过Western blot检测小鼠脊髓组织中TLR4/NF-κB通路蛋白表达水平。结果与正常对照组比较,EAE模型组小鼠神经行为学评分升高,并且伴随体质量降低,脊髓组织大范围髓鞘脱失,呈现大量炎性细胞浸润,髓鞘标志蛋白MBP表达下调,TLR4/NF-κB通路蛋白表达水平升高。经GLPS干预后,EAE小鼠神经功能评分降低、体质量上升,髓鞘脱失改善,炎性细胞浸润减少,Foxp3的表达增加,IL-17的含量降低,TLR4/NF-κB通路蛋白表达降低。结论GLPS能够抑制MS动物模型EAE的发病进程,其作用机制可能是通过抑制TLR4/NF-κB炎症通路改善髓鞘脱失,从而发挥对中枢神经系统的保护作用。Aim To explore the protective effects of ganoderma lucidum polysaccharides(GLPS)on experimental autoimmune encephalomyelitis(EAE),an animal model of multiple sclerosis(MS)and the underlying mechanism.Methods Thirty C57BL/6 mice were randomly divided into three groups:normal control group,EAE model group and GLPS group(5 mg·kg^(-1)).The EAE mouse model was established by injection of mycobacterium tuberculosis and MOG35-55,and the effect of GLPS on body weight and neurological dysfunction score of EAE mice were observed.The pathological changes of myelin sheath in frozen sections of spinal cord of mice were observed by solid blue staining and HE staining,and the expression of MBP was observed by immunofluorescence.The contents of IL-17 and Foxp3 in spleen and lymph nodes of mice were detected by flow cytometry.The expression of TLR4/NF-κB pathway protein in spinal cord tissue of mice was detected by Western blot.Results Compared with the normal control group,the neurological function scores of EAE mice group increased,accompanied with weight loss.Large-scale demyelination of spinal cord tissue and a massive inflammatory cell infiltration were found.Myelin marker protein MBP expression was reduced.The expression of TLR4/NF-κB protein increased.After GLPS intervention,the neurological function score of EAE mice decreased,body weight increased,demyelination was improved,and inflammatory cell infiltration decreased.Meanwhile the expression of Foxp3 increased,the content of IL-17 decreased and the expression of TLR4/NF-κB was inhibited.Conclusions GLPS can inhibit the pathogenesis of EAE in EAE mice,which may be related to the improvement of the demyelination by inhibiting the TLR4/NF-κB inflammatory pathway,thereby exerting a protective effect on the central nervous system.

关 键 词：灵孢多糖 实验性自身免疫性脑脊髓炎 髓鞘脱失 抗炎 神经保护 TLR4/NF-κB通路 

分 类 号：R-332[医药卫生] R284.1R322.81R392.11R744.3