核因子E2相关因子2对甲基苯丙胺诱导小胶质细胞激活和极化的调控作用研究  

Regulation of methamphetamine-induced activation and polarization of microglia by Nrf2

作　　者：杨根梦 彭艳霞 张鑫杰 侯雨含 许婧 李利华 杜琦 洪仕君 YANG Genmeng;PENG Yanxia;ZHANG Xinjie;HOU Yuhan;XU Jing;LI Lihua;DU Qi;HONG Shijun(NHC Key Laboratory of Drug Addiction Medicine,School of Forensic Medicine,Kunming Medical University,Kunming 650500,China)

机构地区：[1]国家卫健委毒品依赖和戒治重点实验室/昆明医科大学法医学院,昆明650500

出　　处：《中国比较医学杂志》2024年第12期1-7,共7页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金(82202081);省科技厅基础研究项目(202301AT070269,202301AU070216);省科技厅-昆医联合专项(202301AY070001-234,202201AY070001-021)。

摘　　要：目的探讨核因子E2相关因子2(Nrf2)对甲基苯丙胺(METH)诱导小胶质细胞激活和极化的调控作用。方法体外实验以小鼠小胶质(BV2)细胞和人小胶质(HMC3)细胞为研究对象;体内实验以野生型小鼠和Nrf2基因敲除鼠为研究对象,分别建立体内外METH诱导的毒性模型,应用免疫荧光(IF)和蛋白免疫印迹检测各组细胞和小鼠脑组织内小胶质细胞激活和极化的表达情况。结果给予METH后,BV2和HMC3细胞表达小胶质细胞M1表型标志蛋白诱导型一氧化氮合酶(iNOS)的荧光水平显著升高(P<0.001),而表达小胶质细胞M2表型标志蛋白精氨酸酶1(Arg1)的荧光水平明显降低(P<0.05,P<0.01);小鼠皮质内小胶质细胞明显被激活,皮质内离子钙结合衔接分子1(IBA1)和iNOS的表达水平明显升高(P<0.001,P<0.05),Arg1的表达水平降低(P<0.01)。与野生型METH组相比,Nrf2基因敲除后给予METH发现小胶质细胞激活的数量明显增多(P<0.01),同时IBA1和iNOS的表达水平有所升高(P<0.001,P<0.01),Arg1的表达水平有所降低(P<0.01)。结论Nrf2对METH诱导小胶质细胞激活和极化具有重要的调控作用,Nrf2有望成为治疗METH诱导神经炎症的潜在干预靶点。Objective To investigate the regulatory role of nuclear factor erythroid 2-related factor 2(Nrf2)in the activation and polarization of microglia induced by methamphetamine(METH).Methods BV2 and HMC3 cells were studied in vitro and wild-type mice and Nrf2-knockout mice were studied in vivo.In vivo and in vitro toxicity models induced by METH were established,respectively.The activation and polarization of microglia in each group were examined using immunofluorescence and Western blot,respectively.Results METH treatment significantly increased the fluorescence level of inducible nitric oxide synthase(iNOS)in BV2 and HMC3 cells(P<0.001),and significantly decreased the fluorescence level of Arginasel(Argl)(P<0.05,P<0.01).METH exposure activated microglia in the cortex,increased expression levels of ionized calcium binding adaptor molecule-1(IBA1)and iNOS(P<0.001,P<0.05),and decreased the expression of Arg1(P<0.01).The number of activated microglia was significantly increased after Nrf2 gene knockout(P<0.O1),compared with the WT METH group,while the expression levels of IBA1 and iNOS were also increased(P<0.001,P<0.01)and the expression level of Argl was decreased(P<0.01).Conclusions Nrf2 plays an important role in regulating the activation and polarization of microglia induced by METH.Nrf2 may thus be a potential target for the treatment of neuroinflammation induced by METH.

关 键 词：甲基苯丙胺 小胶质细胞 核因子E2相关因子2 神经毒性作用 

分 类 号：R-33[医药卫生]