大麻素2型受体拮抗剂对激活态BV2小胶质细胞的免疫调节作用

目的观察大麻素2型受体(cannabinoid 2 receptor, CB2R)拮抗剂对BV2小胶质细胞免疫调节功能的影响。方法 使用适量浓度的IFN-γ刺激激活BV2小胶质细胞,建立模拟EAE炎性环境的细胞模型,比较静息态BV2细胞组、激活态BV2细胞组和大麻素2型受体拮抗剂SR144528A(SR2)干预组CB2R mRNA和蛋白的表达情况,用ELISA方法检测细胞因子和趋化因子的浓度,MTT法检测细胞增殖率。结果 IFN-γ激活的BV2小胶质细胞CB2R mRNA和蛋白的表达均高于静息组(P<0.05)；使用SR2干预激活的BV2小胶质细胞,可降低其CB2R mRNA和蛋白的表达(P<0.05)；SR2可显著上调激活态BV2细胞致炎因子IFN-γ、IL-17、IL-6和TNF-α的水平,促进BV2小胶质细胞增殖和NO的释放(P<0.05),同时显著下调IL-4和MCP-1的浓度,对IL-1β、IL-10、CX3CL1无调节作用。结论CB2R参与了小胶质细胞介导的炎症反应,CB2R在调节Th1/Th17/Th2细胞因子网络平衡中发挥了一定的作用。
Objective To study the effect of cannabinoid 2 receptor(CB2R) antagonist on the activated microglia BV2 cells. Methods Stimulated by IFN-γ, BV2 cells were exposed to CB2R antagonist SR144528A(SR2). The expression of CB2R mRNA and protein were measured by SYBR Green-based Real-Time RT-PCR and Western-blotting, respectively; the levels of cytokines and chemokines were measured with ELISA; and cell proliferation was tested by MTT. ResultsStimulated by IFN-γ, the expression of CB2R in BV2 cells increased significantly(P<0.05). SR2 decreased the expression of CB2R, increased the production of inflammatory cytokines IFN-γ, IL-17, IL-6 and TNF-α as well as NO, promoted cell proliferation, and the production of IL-4 and MCP-1 were decreased; but had no effects on the secretion of IL-1β, IL-10 and CX3CL1. ConclusionCB2R is involved in the inflammatory response mediated by microglia BV2 cells, and CB2R plays a role in the regulation of Th1/Th17/Th2 cytokine network balance