L型Ca^(2+)通道阻断剂通过抑制NLRP3炎症小体激活对MPP+诱导的小胶质细胞炎症模型发挥神经保护作用  

L-type Ca^(2+)channel blocker exerts neuroprotective effect on MPP^(+)-induced microglia inflammation model by inhibiting NLRP3 inflammasome activation

作　　者：刘思昆 陈曦 许苗菁 赵振强[1] 周畅 许叶 王埮[2] Liu Sikun;Chen Xi;Xu Miaoqing;Zhao Zhenqiang;Zhou Chang;Xu Ye;Wang Tan(Key Laboratory of Tropical Brain Research and Translation,Hainan Medical College,Haikou 571199,China;Hainan Medicine Ward 1,Department of Neurology,The First Affiliated Hospital of the Hospital)

机构地区：[1]海南医学院热带脑科学研究与转化重点实验室,海口571199 [2]海南医学院第一附属医院神经内科一病区

出　　处：《脑与神经疾病杂志》2023年第4期245-251,共7页Journal of Brain and Nervous Diseases

基　　金：国家自然科学基金项目(81760237);海南省临床医学中心建设项目资助(2021);海南省自然科学基金高层次人才项目(821RC1125)。

摘　　要：目的在MPP^(+)诱导的帕金森(PD)细胞炎症模型中,使用L型Ca^(2+)通道阻断剂伊拉地平对BV2小胶质细胞进行干预,观察其炎症反应的影响探讨L型Ca^(2+)通道阻断剂伊拉地平是否可以调控NLRP3炎症小体通路.方法使用PD造模药物MPP^(+)在BV2小胶质细胞上制备PD炎症细胞模型,CCK8法分别检测不同浓度MPP^(+)和伊拉地平处理后对BV2的小胶质细胞具体活性影响,选择合适的药物实验浓度.将BV2的小胶质细胞变为4组,分别为空白对照组、MPP^(+)造模组、伊拉地平预处理组和MCC950预处理组(即阳性对照组),采用实时的荧光定量PCR检测BV2小胶质细胞中IL-1β、NLRP3、CASPASE-1、GSDMD、NOX2基因mRNA表达水平;细胞免疫荧光检测BV2小胶质细胞中IL-1β、NLRP3、CASPASE-1、GSDMD、NOX2蛋白表达水平;Western blot检测小胶质细胞IL-1β、NLRP3、CASPASE 1、GSDMD、NOX2蛋白表达水平.结果与空白对照组相比,MPP^(+)(100μmol·L^(-1))能明显降低BV2细胞活性,显著升高IL-1β,NLRP3、CASPASE-1、GSDMDmRNA表达水平(P<0.05).与空白组比较,伊拉地平(5μmol·L^(-1))干预后,减缓了MPP^(+)导致的细胞活性降低(P<0.05),5μmol·L^(-1)伊拉地平可显著降低IL-1β、NLRP3、CASPASE1、GSDMDmRNA表达水平(P<0.05),显著降低MPP^(+)诱导的IL-1β、NLRP3、CASPASE 1、GSDMD蛋白表达水平.结论在PD炎症细胞模型中,L型Ca^(2+)通道阻断剂伊拉地平可能通过NLRP3通路减轻了小胶质细胞炎症因子的表达.Objective In the MPP^(+)-induced Parkinson's disase(PD)cell inflammation model,the L-type Ca^(2+)channel blocker isradipine was used to intervene on BV2 microglia to observe the effect of its inflammatory response to explore whether the L-type Ca^(2+)channel blocker isradipine regulation of the NLRP3 inflammasome pathway.Methods The PD inflammatory cell model was established on BV2 microglia by using the PD modeling drug MPP^(+).The effects of different concentrations of MPP^(+)and isradipine on the activity of BV2 microglia were detected by CCK8 way,and appropriate drug was selected for the experiment.concentration.The BV2 microglia cells were separate into 4 groups,namely blank control group,MPP^(+)modeling group,isradipine pretreatment group and MCC950 pretreatment group(that is,positive control group).PCR to detect the mRNA expression levels of IL-1β,NLRP3,CASPASE-1,GSDMD and NOX2 in BV2 microglia;cellular immunofluorescence to detect IL-1β,NLRP3,CASPASE-1,GSDMD,NOX2 in BV2 microglia protein expression level;Western blot was used to detect the protein expression levels of IL-1β,NLRP3,CASPASE1,GSDMD and NOX2 in microglia.Result Compared with the blank control group,treating with MPP^(+)(100μmol·L^(-1)can significantly reduce the activity of BV2 cells and dramatically increase the mRNA transcription levels·L^(-1)),the decreased cell activity caused by MPPt was slowed down(P<0.05),which also significantly inhibited the transcription and expression levels of IL-1β,NLRP3,CASPASE1,and GSDMD(P<0.05).Conclusion In the PD inflammatory cell model,L-type Ca^(2+)channel blocker isradipine may inhibit the inflammatory factors'expression through the NLRP3 pathway.

关 键 词：伊拉地平 小胶质细胞 NLRP3 帕金森病 

分 类 号：R742.3[医药卫生—神经病学与精神病学]