羟基红花黄色素A减轻双环己酮草酰二腙小鼠髓鞘脱失的机制  

Mechanism by which hydroxysafflor yellow A alleviates demyelination in cuprizone mice

作　　者：陈莹 刘健 梁亚杰 李彦青 宋丽娟 黄建军 尉杰忠[3] 王青[1] 马存根 Chen Ying;Liu Jian;Liang Yajie;Li Yanqing;Song Lijuan;Huang Jianjun;Yu Jiezhong;Wang Qing;Ma Cungen(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,Shanxi Province,China;Department of Neurosurgery,Sinopfarm Tongmei General Hospital/Key Laboratory of Neurological Disease Prevention and Control of Shanxi Provincial Health Commission,Datong 037003,Shanxi Province,China;Institute of Brain Science,Shanxi Datong University/Fifth People’s Hospital of Datong,Datong 037009,Shanxi Province,China)

机构地区：[1]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/神经生物学研究中心,山西省晋中市030619 [2]国药同煤集团总医院神经外科/山西省卫健委神经疾病防治研究重点实验室,山西省大同市037003 [3]山西大同大学脑科学研究所/大同市第五人民医院,山西省大同市037009

出　　处：《中国组织工程研究》2025年第25期5311-5319,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(81903596),项目负责人:王青;山西省卫健委医学科技领军团队(2020TD05),项目负责人:马存根;山西中医药大学学科建设经费(2024XKJS-02),项目负责人:马存根;山西省科技创新人才青年团队项目(202204051001028),项目负责人:宋丽娟;山西省卫健委2022年度中医药科研课题立项计划(2022ZYYC090),项目负责人:马存根;山西中医药大学2022年度科技创新团队(2022TD2006),项目负责人:王青;山西中医药大学2022年度科技创新团队(2022TD2010),项目负责人:宋丽娟;国药同煤总医院横向课题(202209SY01),项目负责人:宋丽娟。

摘　　要：背景:在中枢神经系统脱髓鞘疾病的发生发展过程中,小胶质细胞导致的神经炎症是主要病理特征,因此抑制炎症反应对缓解髓鞘脱失非常重要。羟基红花黄色素A有保护血脑屏障、抑制神经元的凋亡、改善神经功能的作用。目的:探究羟基红花黄色素A抑制双环己酮草酰二腙诱导的小鼠髓鞘脱失的作用机制。方法:①体内实验:将30只健康雄性C57BL/6小鼠随机分为正常组、双环己酮草酰二腙组和羟基红花黄色素A组3组,后2组小鼠喂养含0.2%双环己酮草酰二腙的饲料6周建立脱髓鞘小鼠模型,正常组小鼠喂养正常饲料;在第4周末,给予羟基红花黄色素A组小鼠腹腔注射20 mg/(kg·d)羟基红花黄色素A,正常组和双环己酮草酰二腙组小鼠腹腔注射生理盐水,持续2周。旷场实验、高架十字迷宫实验评价小鼠的行为学变化;黑金染色和髓鞘碱性蛋白、降解髓鞘碱性蛋白免疫荧光染色检测胼胝体髓鞘脱失情况;离子钙结合接头分子1免疫荧光染色和ELISA法分别检测小胶质细胞的活化和炎症因子的表达;Western Blot法检测各组小鼠脑中Toll样受体4、髓样分化因子88、核因子κB p65蛋白的表达水平;②体外实验:采用脂多糖诱导建立BV2小胶质细胞炎症模型。将BV2细胞分为正常组、脂多糖组(1μg/mL)和脂多糖(1μg/mL)+羟基红花黄色素A(25μmol/L)组,采用ELISA法检测细胞上清中肿瘤坏死因子α和白细胞介素6的表达水平。结果与结论:①对比正常组,双环己酮草酰二腙组小鼠焦虑情况严重、自主运动能力异常,胼胝体区髓鞘大量脱失,髓鞘碱性蛋白平均荧光强度显著降低,降解髓鞘碱性蛋白平均荧光强度显著升高,离子钙结合接头分子1阳性小胶质细胞数量增多,脑内白细胞介素1β、肿瘤坏死因子α、白细胞介素6水平升高,Toll样受体4、髓样分化因子88、核因子κB p65的蛋白表达水平明显升高。羟基红花黄色素A治BACKGROUND:In the occurrence and development of demyelinating diseases of the central nervous system,neuroinflammation caused by microglia is the main pathological feature,so inhibiting the inflammatory response is very important to alleviate demyelination.Hydroxysafflor yellow A can protect the bloodbrain barrier,inhibit neuronal apoptosis,and improve neurological function.OBJECTIVE:To explore the mechanism of hydroxysafflor yellow A inhibiting bicyclohexanone oxalyl dihydrazone-induced demyelination in mice.METHODS:(1)In vivo:Thirty healthy male C57BL/6 mice were randomly divided into three groups:normal group,cuprizone group,and hydroxysafflor yellow A group.The mice in the cuprizone group and the hydroxysafflor yellow A group were fed with 0.2%cuprizone diet for 6 weeks to establish mouse models of demyelination.The mice in the normal group were fed with normal diet.At the end of the 4th week,the mice in the hydroxysafflor yellow A group were intraperitoneally injected with hydroxysafflor yellow A 20 mg/kg per day.The mice in the normal and cuprizone groups were intraperitoneally injected with normal saline for 2 weeks.The behavioral changes of mice were evaluated by open field test and elevated plus maze test.The loss of myelin sheath in corpus callosum was detected by black gold staining,myelin basic protein and degraded myelin basic protein immunofluorescence staining.The activation of microglia and the expression of inflammatory factors were detected by Iba-1 immunofluorescence staining and ELISA,respectively.The protein expression levels of Toll-like receptor 4,myeloid differentiation factor 88,and nuclear factorκB p65 in the brain of mice in each group were detected by western blot assay.(2)In vitro experiment:The inflammation model of BV2 microglia was established by lipopolysaccharide induction.BV2 cells were divided into normal group,lipopolysaccharide group(1μg/mL),and lipopolysaccharide(1μg/mL)+hydroxysafflor yellow A(25μmol/L)group.The expression levels of tumor necrosis factorαand interleuki

关 键 词：羟基红花黄色素A 双环己酮草酰二腙 炎症 髓鞘脱失 小胶质细胞 小鼠 

分 类 号：R446[医药卫生—诊断学] R318[医药卫生—临床医学] R744.5