甲基苯丙胺引起BV2小胶质细胞炎性反应:基于Toll样受体-Peli1信号通路的研究  

Methamphetamine induced neuroinflammation in microglial BV2 cells based on the roles of Toll like receptors-Peli1 pathway

作　　者：须怀沙 王易欣[2] 陈旭锋[3] 蒋雷[3] 王军[2] 张久平[4] XU Huaisha;WANG Yixin;CHEN Xufeng;JIANG Lei;WANG Jun;ZHANG Jiuping(Department of Clinical Psychology,Nanjing Drum Tower Hospital,Nanjing 210008;Department of Toxicology y School of Public Health,Nanjing Medical University,Nanjing 211166;Department of Emergency,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029;Child Mental Health Research Center,Nanjing Brain Hospital Affiliated to Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京鼓楼医院医学心理科,江苏南京210008 [2]南京医科大学公共卫生学院卫生毒理系,江苏南京211166 [3]南京医科大学第一附属医院急诊科,江苏南京210029 [4]南京医科大学附属脑科医院儿童心理卫生研究中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2021年第3期324-330,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81673213);江苏省自然科学基金(BK20191349)。

摘　　要：目的:探讨Toll样受体(Toll like receptor,TLR)-E3泛素连接酶Pellino 1(Peli1)介导的炎性通路在甲基苯丙胺(methamphetamine,Meth)引起BV2小胶质细胞炎性反应中的作用。方法:利用Western blot观察Meth作用后Toll样家族中多种TLR的表达及其下游接头蛋白髓样分化因子(myeloid differentiation factor 88,MyD88)、β干扰素TIR结构域衔接蛋白(TIR-domaincontaining adaptor inducing interferon-β,TRIF)水平的改变,同时观察上述接头蛋白下游Peli1蛋白的表达,利用ELISA、实时定量PCR(real time-PCR)及Western blot观察Peli1调节的下游炎性因子及信号通路的改变。结果:Meth作用于BV2细胞后,TLR3、TLR4、TLR7、TLR8、TLR11在特定时间内表达明显上调,同时下游MyD88及TRIF蛋白表达显著增加,其中TRIF具有浓度依赖效应。Meth作用后亦可引起泛素化蛋白Peli1的表达,而利用RNA干扰的方法将Peli1下调后,炎性因子诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、肿瘤坏死因子(tumor necrosis factor,TNF)-α、白细胞介素(interleukin-6,IL)-6表达下降,核因子(nuclear factor,NF)-κB的激活明显缓解。结论:TLR介导的炎性信号在Meth引起BV2细胞炎性反应过程中发挥重要作用。因此,基于TLRs-Peli1靶信号轴的干预可能为Meth神经毒性的干预提供靶点,具有潜在的应用意义。Objective:This study aims to investigate the roles of Toll like receptor(TLR)-Pellino 1(Peli1)axis in methamphetamine(Meth)-mediated microglial inflammation and pathway in BV2 cells. Methods:Western blot was used to observe the expression of Toll like receptors(TLR),the downstream adaptor proteins of myeloid differentiation factor 88(MyD88)and TIR-domain-containing adaptor inducing interferon-β(TRIF). In parallel,the expression of Peli1 protein was detected by Western blot. The downstream inflammatory factors and signal pathway regulated by Peli1 were observed by ELISA,real-time PCR and Western blot. Results:The expression of TLR3,TLR4,TLR7,TLR8 and TLR11 was significantly up-regulated after Meth treatment in BV2 cells. Meanwhile,the expression of MyD88 and TRIF was significantly increased,and TRIF level exhibited a concentration-dependent effect. Meanwhile,Meth obviously induced the expression of ubiquitin protein Peli1. Noteworthily,after Peli1 was down-regulated by RNA interference,the expression of inflammatory factors including inducible nitric oxide synthase(iNOS),tumor necrosis factor(TNF)-α and interleukin(IL)-6 were dramatically decreased,accompanied by the significant attenuation of nuclear factor(NF)-κB activation. Conclusion:The TLRs-mediated inflammatory signal plays an important role in the process of microglial inflammatory response induced by Meth. Therefore,the TLRs-Peli1 signal axis may be a promising target for the intervention of Meth neurotoxicity,which displays significant application.

关 键 词：甲基苯丙胺 TOLL样受体家族 Peli1 神经炎性反应 BV2细胞 

分 类 号：R392.12[医药卫生—免疫学]