法舒地尔通过抑制NLRP3炎症小体激活抑制Aβ_(1-42)诱导的小胶质细胞炎症反应  

Fasudil inhibits Aβ_(1-42)-induced microglial inflammatory response by inhibiting activation of NLRP3 inflammasome

作　　者：郭敏芳[1] 章培军[1] 于婧文[1] 孟涛 李艳花[1] 李娜 李梦迪 李玉璐 宋丽娟 尉杰忠 马存根[1,2] GUO Minfang;ZHANG Peijun;YU Jingwen;MENG Tao;LI Yanhua;LI Na;LI Mengdi;LI Yulu;SONG Lijuan;YU Jiezhong;MA Cungen(Institute of Brain Science,Shanxi Datong University,Datong 037009,China;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 030619,China;Department of Physiology,Shanxi Medical University,Taiyuan 030001,China;Datong Fifth People's Hospital,Datong 037008,China)

机构地区：[1]山西大同大学脑科学研究所,大同037009 [2]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,晋中030619 [3]山西医科大学生理学系,太原030001 [4]大同市第五人民医院,大同037008

出　　处：《中国免疫学杂志》2024年第9期1833-1837,共5页Chinese Journal of Immunology

基　　金：山西省基础研究计划项目(20210302123476,20210302123475,20210302123337);山西省卫生健康委医学科技领军团队项目(2020TD05);国家中医药管理局多发性硬化益气活血重点研究室开放项目(2021-KF-04T);山西中医药大学青年科学家培育项目(2021-PY-QN-09);山西中医药大学2022年度科技创新团队项目(2022TD2010)。

摘　　要：目的:基于NLRP3炎症小体探讨法舒地尔减轻Aβ_(1-42)诱导的BV2小胶质细胞损伤的机制。方法:BV2细胞分为:正常对照组、Aβ刺激组、Aβ+法舒地尔联合干预组、Aβ+MCC950(NLRP3抑制剂)联合干预组。显微镜下观察细胞形态;CCK8测定细胞活性;Griess测定NO释放量;免疫荧光染色检测NLRP3、caspase 1和IL-18表达;Western blot检测NLRP3、ASC、caspase 1、IL-1β和IL-18表达。结果:与正常对照组比较,Aβ_(1-42)刺激组BV2细胞激活,呈阿米巴样形态,活性下降,NO释放量增加,NLRP3、ASC、caspase 1、IL-1β和IL-18表达增加。法舒地尔干预和MCC950干预均可改善Aβ_(1-42)诱导的BV2细胞损伤,使细胞形态趋于正常,细胞活性有所增加,NO释放量降低,同时下调NLRP3、ASC、caspase 1、IL-1β和IL-18表达,两组间差异无统计学意义。结论:法舒地尔可能通过抑制NLRP3炎症小体激活减轻Aβ_(1-42)诱导的BV2细胞损伤和炎症反应。Objective:To explore mechanism of Fasudil reducing Aβ_(1-42)induced BV2 cell injury based on NLRP3 inflammasome.Methods:BV2 cells were divided into:normal control group,Aβstimulation group,Aβ+Fasudil intervention group,Aβ+MCC950(NLRP3 inhibitor)intervention group.Cell morphology was observed under microscope.Cell activity was determined of by CCK8.NO release was measured by Griess.NLRP3,caspase 1 and IL-18 expressions were detected by immunofluorescence staining.NLRP3,ASC,caspase 1,IL-1βand IL-18 expressions were detected by Western blot.Results:Compared with normal control group,BV2 cells in Aβstimulation group were activated and showed amoeba-like shape,cell activity was decreased,NO production was increased,NLRP3,ASC,caspase 1,IL-1βand IL-18 expressions were increased.Fasudil intervention and MCC950 intervention inhibited cell injury induced by Aβ_(1-42)in which BV2 cell morphology tended to be normal,cell activity was increased,while production of NO was reduced,and NLRP3,ASC,caspase 1,IL-1βand IL-18 expressions were down-regulated,there was no significant difference between Fasudil intervention group and MCC950 intervention group.Conclusion:Fasudil may alleviate Aβ_(1-42)induced BV2 cell injury and inflammatory reaction by inhibiting NLRP3 inflammasome activation.

关 键 词：法舒地尔 小胶质细胞 NLRP3炎症小体 炎症反应 

分 类 号：R392.1[医药卫生—免疫学]