摘要
目的:基于NLRP3炎症小体探讨法舒地尔减轻Aβ1-42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,Aβ1-42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Aβ1-42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Aβ1-42诱导的BV2细胞损伤和炎症反应.
Abstract
Objective:To explore mechanism of Fasudil reducing Aβ1-42 induced BV2 cell injury based on NLRP3 inflamma-some.Methods:BV2 cells were divided into:normal control group,Aβ stimulation 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-1β and IL-18 expressions were detected by Western blot.Results:Compared with normal control group,BV2 cells in Aβ stimulation group were activated and showed amoeba-like shape,cell activity was decreased,NO production was increased,NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were increased.Fasudil intervention and MCC950 intervention inhibited cell injury induced by Aβ1-42 in which BV2 cell morphology tended to be normal,cell activity was increased,while produc-tion of NO was reduced,and NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were down-regulated,there was no significant difference between Fasudil intervention group and MCC950 intervention group.Conclusion:Fasudil may alleviate Aβ1-42 induced BV2 cell injury and inflammatory reaction by inhibiting NLRP3 inflammasome activation.
基金项目
山西省基础研究计划项目(20210302123476)
山西省基础研究计划项目(20210302123475)
山西省基础研究计划项目(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)
国家科技期刊平台
NSTL
万方数据