AM1241抑制ADPβS诱发培养大鼠脊髓背角小胶质细胞P2Y_(12)和P2Y_(13)受体表达及炎症因子释放  

AM1241 inhibits the expression of P2Y_(12)/P2Y_(13) receptors and inflammatory cytokines release from ADPβS-activated dorsal spinal cord microglia cells

作　　者：乐明霞 徐陶[1] 周瑞[1] 黄杜娟[1] 杨俊娜[1] 何丽[1] 刘晓红[1] 曾俊伟[1] 

机构地区：[1]遵义医学院生理学教研室贵州省麻醉与器官功能保护重点实验室,遵义563000

出　　处：《神经解剖学杂志》2018年第1期34-40,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(1460266;31640040);教育部新世纪优秀人才计划(NCET-13-1070);贵州省科技厅课题[黔科合J字(2010)2264号]

摘　　要：目的:离体条件下,观察大麻素CB2受体激动剂AM1241对嘌呤P2Y受体激动剂ADPβS诱发的脊髓背角小胶质细胞P2Y_(12)和P2Y_(13)受体mRNA表达以及炎症因子IL-1β、IL-6和TNF-α释放的影响。方法:培养纯化新生SD大鼠(<3 d)脊髓背角小胶质细胞,荧光定量PCR技术检测AM1241(10^(-5)mol/L,1 h)对ADPβS(10^(-5)mol/L,3 h)诱发的背角小胶质细胞P2Y_(12)和P2Y_(13)受体mRNA表达的影响;ELISA技术检测AM1241(10^(-5)mol/L,1 h)对ADPβS(10^(-5)mol/L,3 h)诱发的小胶质细胞IL-1β、IL-6和TNF-α释放的影响。结果:与正常对照组相比,ADPβS(10^(-5)mol/L,3 h)可以刺激脊髓背角小胶质细胞P2Y_(12)和P2Y_(13)受体在mRNA水平表达上调(P<0.05),IL-1β、IL-6和TNF-α释放相应增加(P<0.05)。AM1241(10^(-5)mol/L,1 h)几乎完全阻断ADPβS刺激小胶质细胞P2Y_(12)和P2Y_(13)受体mRNA表达上调以及IL-1β、IL-6和TNF-α释放的效应(P<0.05);AM1241的这种抑制效应可以被CB2受体拮抗剂AM630(10^(-5)mol/L,1 h)反转(P<0.05)。结论:大麻素CB2受体激活可以抑制ADPβS诱发的脊髓背角小胶质细胞P2Y_(12)和P2Y_(13)受体mRNA水平表达上调以及IL-1β、IL-6和TNF-α释放。Objective： To investigate the effect of AM1241 （selective CB2 receptor agonist） on ADPβS-evoked P2Y12 and P2Y13 receptors mRNA expression and inflammatory cytokines release from cultured dorsal spinal cord microglia cells. Methods： A cell culture model of SD rat dorsal spinal cord micmglia was used. The influence of AM1241 （10^-5 mol/L, 1 h）on ADPβS（ 10^-5 mol/L, 3 h）-evoked P2Y12 and P2Y13 receptors mRNA expression were observed by using real time fluorescence quantitative PCR （RTFQ-PCR）. The influence of AM1241 （ 10^-5 mol/L, 1 h） on ADPβS（ 10^-5 mol/L, 3 h ）-evoked IL-1β, IL-6 and TNF-α release were measured by ELISA assay. Results： Compared with the control, stimulation of microglia cells with ADPβS（ 10^-5 mol/L, 3 h）led to a robust increase of P2YI2 and P2Y13 receptors mRNA （P 〈0. 05 ）. ADPβS （10^-5 tool/L, 3 h）-induced IL-1β, IL-6 and TNF-α release were also higher than that of control groups （P 〈0. 05）. ADPβS-indueed the increased expression of P2Y12 and P2Y13 receptors mRNA was significantly inhibited after pretreatment with AM1241 at 10^-5 mol/L for lh （P 〈0.05）. Similarly, ADPβS-induced IL-1β, IL-6 and TNF-α release were obvious inhibited after pretreatment with AM1241 at 10^-5 mol/L for 1 h （P 〈 0.05 ）. Furthermore, the inhibitory effects of AM1241 on ADP13S-evoked P2Y12 and P2Y13 receptors mRNA expression and inflammatory cytokines release were significantly reversed after administration of CB2 receptor antagonist AM630 at 10^-5 mol/L for 1 h （P 〈 0.05）. Conclusion： CB2 receptor activation significantly suppressed ADPβS-evoked P2Y12 and P2Y13 receptors mRNA expression and inflammatory cytokines release from cultured rat dorsal spinal cord microglia cells.

关 键 词：P2Y12受体 P2Y13受体 小胶质细胞 IL-1Β IL-6 TNF-α 大鼠 

分 类 号：R741[医药卫生—神经病学与精神病学]