细叶远志皂苷通过抑制炎症与氧化应激减轻CPZ诱导的小鼠髓鞘脱失  

Tenuifolin reduces CPZ-induced demyelination in mice by inhibiting inflammation and oxidative stress

作　　者：李彦青 杨智超 王翰斌 杨立志 宋丽娟 肖保国[3] 王青 马存根 LI Yan-qing;YANG Zhi-chao;WANG Han-bin;YANG Li-zhi;SONG Li-juan;XIAO Bao-guo;WANG Qing;MA Cun-gen(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,030619,China;Shanxi Guorun Pharmaceutical Co.,Ltd,Datong,038100,China;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：《时珍国医国药》2023年第11期2599-2605,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81903596);山西中医药大学博士来晋工作启动基金(2021BKS01);山西中医药大学博士启动基金(2022BK15);山西中医药大学培育计划项目(2021PY-JC-15);山西科技厅山西省基础研究青年项目(20210302123312);横向课题(2022HX-GR-01)。

摘　　要：目的探究细叶远志皂苷(TEN)治疗双环己酮草酰二腙(CPZ)诱导的脱髓鞘小鼠模型的作用机制。方法实验将24只C57BL/6雄性小鼠随机分为对照组、CPZ模型组和TEN用药组,每组8只。对照组小鼠给予正常饲料,模型组和用药组喂养含0.2%(质量分数)CPZ饲料6周,自造模第5周开始,用药组给予TEN灌胃治疗2周。采用旷场实验、爬杆实验与平衡木实验评价小鼠的运动协调功能;使用免疫荧光染色检测各组小鼠脑中MBP、dMBP、Iba1、NF-κB p65的平均荧光强度;通过ELISA法检测各组小鼠血清中IL-1β、TNF-α的含量;Western blot法检测各组小鼠脑中MBP、Nrf2、HO-1的蛋白表达水平。结果与对照组相比,CPZ模型组小鼠运动协调能力明显下降,脑组织中髓鞘标志蛋白MBP显著降低、Iba1阳性小胶质细胞数量及NF-κB表达增多,血清中IL-1β、TNF-α水平升高,Nrf2、HO-1的蛋白表达水平升高,通过TEN干预,改善了小鼠运动功能障碍,胼胝体髓鞘含量升高,小胶质细胞活化及炎症反应得到抑制,血清中IL-1β、TNF-α水平降低,脑组织匀浆中Nrf2、HO-1的蛋白表达水平显著升高。结论TEN能够改善CPZ诱导的脱髓鞘小鼠的运动功能障碍,抑制小胶质细胞活化介导的炎症反应,减轻小鼠脑内的氧化应激,从而缓解髓鞘脱失。Objective To explore the mechanism of tenuifolin(TEN)in the treatment of demyelinating mouse model induced by cuprizone(CPZ).Methods Twenty-four C57BL/6 male mice were randomly divided into control group,CPZ group and treatment group,with 8 mice in each group.The mice in the control group were fed with normal diet,the mice in the model group and the treatment group were fed with 0.2%(mass fraction)CPZ diet for 6 weeks.From the 5 th week of modeling,the mice in the treatment group were given TEN for 2 weeks.The motor coordination function of mice were evaluated by open field test,pole test and beam-walking test.The average fluorescence intensity of MBP,dMBP,Iba1 and NF-κB p65 in the brain of each group was detected by immunofluorescence staining.The contents of IL-1β and TNF-αin serum of mice in each group were detected by ELISA.The protein expression levels of MBP,Nrf2 and HO-1 in the brain of each group were detected by Western blot.Results Compared with the control group,the CPZ group decreased motor coordination within 6 weeks.The content of MBP in corpus callosum decreased,the number of microglia and the expression of NF-κB increased,the levels of IL-1β and TNF-α in serum increased,and the protein expression levels of Nrf2 and HO-1 in brain homogenate increased.TEN intervention improved motor coordination in mice,increased myelin content and inhibited microglial activation and inflammatory response in corpus callosum.The levels of IL-1β and TNF-α in serum were decreased,and the protein expression levels of Nrf2 and HO-1 in brain homogenate were futher increased.Conclusion TEN can ameliorate the motor dysfunction in the CPZ-induced demyelinating mice,inhibit microglia activation-mediated inflammatory response,and reduce oxidative stress in the brain of mice via the activation of Nrf2 and HO-1 signaling molecules,thereby alleviating the demyelination.

关 键 词：细叶远志皂苷 双环己酮草酰二腙 炎症 氧化应激 髓鞘脱失 

分 类 号：R285.5[医药卫生—中药学] R329.21[医药卫生—中医学]