银杏内酯B口服对cuprizone诱导的脱髓鞘小鼠的髓鞘保护  

Protective effects of oral ginkgolide B on demyelination induced by cuprizone in mice

作　　者：俞明杨 赵梦茹 田童 张倩霞 张雨萌 杨颖博[1,2,3] 肖保国 肖伟[1,2] YU Mingyang;ZHAO Mengru;TIAN Tong;ZHANG Qianxia;ZHANG Yumeng;YANG Yingbo;XIAO Baoguo;XIAO Wei(不详;Institute of Chinese Materia Medica,Shanghai University of Chinese Medicine,Shanghai 200120,China;Jiangsu Kanion Pharmaceutical Co.,Ltd.,Lianyungang 222001,China)

机构地区：[1]上海中医药大学中药研究所,200120 [2]江苏康缘药业股份有限公司,222001 [3]上海图锋医药科技有限公司,200120 [4]上海复旦大学神经病学研究所,200040

出　　处：《中国神经免疫学和神经病学杂志》2021年第5期386-391,共6页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家重点研发计划资助项目(2019YFC1711000)。

摘　　要：目的探讨口服银杏内酯B(GB)对双环己酮草酰二腙(cuprizone,CPZ)诱导的脱髓鞘小鼠的保护作用。方法选择C57BL/6雄性小鼠24只,随机将小鼠分为正常组、CPZ模型组和GB干预组,每组8只。采用连续喂食6周含0.2%(质量分数)CPZ的饲料方法制备脱髓鞘小鼠模型。正常组小鼠喂食正常饲料。从造模第3周开始,GB干预组小鼠给予GB灌胃,连续4周,正常组和CPZ模型组给予等量0.5%(质量分数)CMC-Na溶剂灌胃。采用高架十字迷宫、强迫游泳和T迷宫分别评估各组小鼠的焦虑、抑郁和认知行为。使用LFB染色检测各组小鼠脑髓鞘脱失情况,使用Western blot和ELISA检测各组小鼠脑组织匀浆中钙离子结合衔接分子1(Iba-1)蛋白表达和肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和IL-1β水平。体外实验采用LPS处理BV-2细胞建立体外小胶质细胞炎症模型,分为正常组细胞、模型组细胞和GB干预组细胞,采用Griess法和ELISA检测细胞上清液中NO、TNF-α、IL-6和IL-1β水平。结果(1)动物实验:与正常组相比,CPZ模型组小鼠出现焦虑、抑郁和认知障碍,脑胼胝体髓鞘含量降低(均P<0.05),脑皮层、胼胝体和纹状体的小胶质细胞数量增多(均P<0.05),脑组织匀浆中Iba-1蛋白及TNF-α、IL-6和IL-1β水平升高(均P<0.05);与CPZ模型组相比,GB可改善焦虑、抑郁,GB干预组小鼠胼胝体髓鞘含量升高(均P<0.05),皮层、胼胝体和纹状体的小胶质细胞数量减少(均P<0.05),脑组织匀浆Iba-1、TNF-α、IL-6和IL-1β水平降低(均P<0.05)。(2)细胞实验:经LPS刺激的BV2细胞上清液NO、TNF-α、IL-6和IL-1β水平高于对照组(均P<0.05),而经GB处理可有效降低NO、TNF-α、IL-6和IL-1β的含量(均P<0.05)。结论口服GB可改善CPZ诱导的脱髓鞘小鼠的行为异常,抑制小胶质细胞活化介导的炎性反应和氧化应激,具有髓鞘保护作用。Objective To investigate the protective effect of oral ginkgolide B(GB)on demyelinating mice induced by cuprizone.Methods Twenty-four male C57BL/6 mice were selected and randomly divided into a normal group,a CPZ model group,and a GB intervention group,with 8 mice in each group.A demyelination mouse model was prepared by feeding continuously for 6 weeks with 0.2%CPZ.Mice in the normal group were fed normal diet.From the 3rd week of modeling,the mice in the GB intervention group were given GB gavage for 4 consecutive weeks,while the normal group and the CPZ model group were given 0.5%CMC-Na solvent gavage.The elevated plus maze,forced swimming and T-maze were used to evaluate the anxiety,depression and cognitive behavior of mice in each group.LFB staining was used to detect demyelination in the brain of mice in each group.Western blot and ELISA were used to detect the protein expression of Iba-1 and the levels of TNF-α,IL-6 and IL-1βin the brain tissue homogenate of mice in each group.In vitro experiment,BV-2 cells were treated with LPS to establish microglial inflammatory model,which were divided into a normal group,a model group and a GB intervention group.The levels of NO,TNF-α,IL-6 and IL-1βin cell supernatant were detected by Griess method and ELISA.Results(1)Animal experiments:compared with the normal group,mice in the model group showed anxiety,depression and cognitive impairment,and the content of myelin sheath in the corpus callosum was reduced(all P<0.05).The number of microglia in the cortex,corpus callosum and striatum was increased(all P<0.05).The expression of Iba-1 protein and the levels of TNF-α,IL-6 and IL-1βin the homogenate of brain tissue were increased(all P<0.05).Compared with the CPZ model group,GB improved anxiety and depression,and the content of myelin sheath in the corpus callosum of mice in the GB intervention group increased(all P<0.05).The number of microglia was decreased in the cortex,corpus callosum and striatum(all P<0.05).The levels of Iba-1,TNF-α,IL-6 and IL-1βin brain t

关 键 词：银杏内酯 双环己酮草酰二腙 脱髓鞘疾病 小胶质细胞 神经炎症 

分 类 号：R744.5[医药卫生—神经病学与精神病学]