藁本内酯通过AIM2/caspase-1信号通路抑制炎症反应改善远端大脑中动脉闭塞小鼠髓鞘脱失  

Ligustilide improves demyelination of dMCAO mouse model by inhibiting inflammation through AIM2/caspase-1 signaling pathway

作　　者：梁亚杰 刘健 陈莹 张紫薇 蒲萌 汤亿彬 张海飞[2] 宋国斌[2] 马存根 王青 LIANG Ya-jie;LIU Jian;CHEN Ying;ZHANG Zi-wei;PU Meng;TANG Yi-bin;ZHANG Hai-fei;SONG Guo-bin;MA Cun-gen;WANG Qing(The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis,State Administration of Traditional Chinese Medicine/Neurobiology Research Center,Shanxi University of Chinese Medicine,Jinzhong Shanxi 030619,China;Institute of Brain Science/Dept of Neurology of First Affiliated Hospital,Shanxi Datong Universiy,Datong Shanxi 037009,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,山西晋中030619 [2]山西大同大学脑科学研究所/附属第一医院神经科,山西大同037009

出　　处：《中国药理学通报》2025年第5期851-860,共10页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81903596);山西省中医药管理局中医药创新团队(No zyytd2024039);山西省回国留学人员科研资助项目(No 2022-165);山西省卫健委医学科技领军团队项目(No 2020TD05);国家中医药管理局科研课题(No 2023ZYYDA2038,2023ZYYB040)。

摘　　要：目的探讨藁本内酯(ligustilide,LIG)通过AIM2/caspase-1信号通路抑制小鼠远端大脑中动脉闭塞(distal middle cerebral artery occlusion,dMCAO)模型的炎症反应而改善髓鞘脱失的机制研究。方法将30只C57BL/6N雄性小鼠随机分为3组:假手术组(Sham组,n=10)、模型组(dMCAO组,n=10)和治疗组(LIG组,n=10)。dMCAO组和LIG组通过电凝法复制dMCAO小鼠模型,小鼠在清醒后进行Longa评分,评分2~3分为造模成功,激光散斑血流成像系统监测小鼠dMCAO术后大脑血流改变状况;造模1 h后LIG组腹腔注射LIG(30 mg·kg^(-1)·d^(-1)),Sham组和dMCAO组注射等量生理盐水,累计1周至实验结束。TTC染色观察小鼠大脑的损伤情况;采用疲劳转棒实验、旷场实验评价小鼠的运动功能以及焦虑程度;后取小鼠脑组织进行黑金染色比较各组髓鞘变化情况;采用免疫荧光染色检测各组小鼠脑内CC区、CPU区和CX区的MBP、IBA1、GFAP的平均荧光强度;采用ELISA法检测小鼠脑组织蛋白中TNF-α、IL-6、IL-1β、IL-17A和BDNF的含量;采用Western blot法检测各组AIM2、caspase-1、ASC的蛋白表达水平。结果与dMCAO组比较,LIG组小鼠梗死面积减少,行为学明显改善,且髓鞘脱失减少。脑组织中CC区、CPU区和CX区中MBP蛋白表达水平升高(P<0.05);CX区的IBA1的蛋白表达水平降低(P<0.01),CC区、CPU区和CX区中GFAP的表达水平升高(P<0.01)。ELISA结果显示,用药后TNF-α(P<0.01)、IL-6(P<0.01)、IL-1β(P<0.05)、IL-17A(P<0.01)水平明显降低,BDNF的表达水平增高(P<0.05),且用药后小鼠脑内AIM2、caspase-1、ASC的蛋白表达水平降低(P<0.01)。结论LIG对dMCAO小鼠模型的髓鞘脱失具有保护作用,其机制可能与抑制AIM2/caspase-1信号通路而抑制了炎症反应、促进星性胶质细胞表达BDNF有关。Aim To explore the mechanism of ligustilide(LIG)improving demyelination by inhibiting inflammatory response in mice with distal middle cerebral artery occlusion(dMCAO)through AIM2/caspase-1 signal pathway.Methods Thirty C57BL/6N male mice were randomly divided into three groups:sham operation group(Sham group,n=10),model group(dMCAO group,n=10)and treatment group(LIG group,n=10).The dMCAO mouse model was established by electrocoagulation in dMCAO group and LIG group.The mice were scored by Longa after waking up,and the changes of cerebral blood flow were monitored by laser speckle blood flow imaging system after dMCAO.One hour after modeling,LIG(30 mg·kg^(-1)·d^(-1))was injected intraperitoneally in LIG group,and the same amount of normal saline was injected in sham group and dMCAO group for one week until the end of the experiment.The mice in each group were stained with TTC,and the brain injury was observed pathologically.Fatigue turning bar test and open field test were used to evaluate the motor function and anxiety degree of mice,and then the brain tissues of mice were taken for black gold staining to compare the changes of myelin sheath in each group.Immunofluorescence staining was used to detect the average fluorescence intensity of MBP,IBA1 and GFAP in CC,CPU and CX regions of mouse brains.ELISAwas used to detect the contents of TNF-α,IL-6,IL-1β,IL-17A and BDNF in brain tissue proteins of mice.Western blotting was used to detect the protein expressions of AIM2,caspase-1 and ASC-in each group.Results Compared with the dMCAO group,the infarct area was reduced,the behavior was significantly improved and the demyelination was reduced in the LIG group.The expression of MBP protein in CC,CPU and CX regions increased(P<0.05),the expression of IBA1 in CX decreased(P<0.01),and the expression of GFAP increased in CC,CPU and CX regions(P<0.01).The results of ELISA showed that the levels ofTNF-α(P<0.01),IL-6(P<0.01),IL-1β(P<0.05)and IL-17A(P<0.01)significantly decreased,while the expression of BDNF increased(P<

关 键 词：藁本内酯 脱髓鞘 炎性反应 dMCAO 小胶质细胞 AIM2/caspase-1信号通路 

分 类 号：R-332[医药卫生] R284.1R322.81R364.5R743.31R744.5