摘要
目的:探讨Toll样受体(Toll like receptor,TLR)/核因子(nuclear factor,NF)-κB信号通路在甲基苯丙胺(methamphetamine,METH)引起原代小胶质细胞激活及炎性因子释放的作用。方法:分离培养原代小胶质细胞,免疫荧光法鉴定原代小胶质细胞纯度。细胞予METH(300μmol/L)染毒0 min、15 min、30 min、1 h、3 h、6 h、12 h和24 h,蛋白免疫印迹检测原代小胶质细胞的激活情况及其NF-κB通路激活情况,real-time PCR检测炎性因子[肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、白介素-6(interleukin-6,IL-6)]和TLR1~9、11 m RNA的表达。结果:分离出的原代小胶质细胞纯度达到95%以上。METH暴露后原代小胶质细胞激活,其激活标志物IBA-1水平增高,NF-κB通路被激活,炎性因子(TNF-α、IL-1β、IL-6)和TLR2、4~5、7~9、11的mRNA水平明显升高,TLR1的mRNA水平明显降低,差异具有统计学意义(P <0.05)。结论:METH可通过调节TLR/NF-κB信号通路激活原代小胶质细胞并促进其释放炎性因子,因而TLR可作为METH神经炎性反应干预的潜在靶点,具有一定的治疗意义。
Objective:To investigate the role of Toll-like receptor(TLR)/NF-κB signaling pathway in methamphetamine(METH)-induced primary microglial activation and inflammatory reaction. Methods:Primary microglial cells were isolated and cultured,then the purity of primary microglia was detected by immunofluorescence. Cells were treated with METH(300 μmol/L)for 0 min,15 min,30 min,1 h,3 h,6 h,12 h,and 24 h,and the activations of primary microglia and NF-κB pathway were detected by Western blot. The inflammatory factors(TNF-α,IL-1β,and IL-6)and TLR1-9,11 mRNA levels were detected by real-time PCR. Results:The purity of primary microglial cells was above 95%. After exposure to METH,the primary microglia activation marker IBA-1 was increased and the NF-κB pathway was activated. In addition,the mRNA levels of inflammatory factors(TNF-α,IL-1β,IL-6)and TLR2,4-5,7-9,11 were significantly increased(P < 0.05),while the level of TLR1 mRNA was significantly reduced. Conclusion:METH can activate primary microglial cells and promote the release of inflammatory factors by regulating the TLR/NF-κB signaling pathway. Therefore,TLR can be potential targets for METH neuritis response intervention with certain therapeutic significance.
作者
肖逸
王胜婵
王易欣
臧松松
张炜
XIAO Yi;WANG Shengchan;WANG Yixin;ZANG Songsong;ZHANG Wei(Department of Urology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029;Department of Urology,Sir Run Run Hospital,Nanjing Medical University,Nanjing 211100;Department of Urology,Geriatric Hospital of Nanjing Medical University,Nanjing 210024;Department of Toxicology,School of Public Health,Nanjing Medical University,Nanjing 211166,China)
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2021年第7期970-975,共6页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81673213)。
