嘌呤P2Y受体激动剂ADPβS对脊髓来源小胶质细胞产生IL-1β、IL-6和TNF-α的刺激作用

P2Y_(12)/ P2Y_(13) receptor are involved in ADPβS- stimulated production of IL-1β,IL-6 and TNF-α from cultured spinal microglia cells

目的:观察嘌呤P2Y受体激动剂ADPβS对脊髓背角来源小胶质细胞炎症因子IL-1β、IL-6和TNF-α表达和释放的影响。方法:培养新生SD大鼠脊髓背角小胶质细胞,PCR检测ADPβS作用下小胶质细胞IL-1β、IL-6和TNF-α在mRNA水平表达变化;ELISA检测ADPβS作用下小胶质细胞IL-1β、IL-6和TNF-α释放变化。结果:与正常对照组相比,ADPβS(10μmol/L)可以刺激脊髓背角小胶质细胞Iba-1、IL-1β、IL-6和TNF-αmRNA表达上调。P2Y_(12)受体拮抗剂MRS2395和P2Y_(13)受体拮抗剂MRS2211部分阻断ADPβS刺激小胶质细胞IL-1β、IL-6和TNF-αmRNA表达上调,MRS2395和MRS2211联合预处理几乎全部阻断ADPβS刺激小胶质细胞Iba-1、IL-1β、IL-6和TNF-αmRNA表达上调。结论:P2Y_(12)/P2Y_(13)受体激活可以促进脊髓背角小胶质细胞激活和IL-1β、IL-6和TNF-αmRNA表达上调及释放。

Objective： To explore whether ADPβS has an effect on the expression and release of IL-1β、IL-6 and TNF-α from cultured spinal cord microglia. Methods： A cell culture model of SD rat spinal cord mieroglia was used. The expression of Iba-1, IL-1β、IL-6 and TNF-α mRNA in cultured dorsal spinal cord microglia were observed by using realtime fluorescence quantitative PCR （RT-PCR）. ADPβS-induced IL-1β、IL-6 and TNF-α release from microglia cells were measured by ELISA assay. Results： Compared with the control, the expression of Iba-1, IL-1β、IL-6 and TNF-α at the mRNA level were obvious increased after ADPβS adiministration. ADPβS-induced IL-1β、IL-6 and TNF-α release were also higher than that of control groups. ADPβS-evoked the increased gene expression of Iba-1,IL-1β、IL-6 and TNF-α was partly inhibited by P2Y12 receptor antagonist MRS2395 and P2Y13 receptor antagonist MRS2211. Similarly, ADPβS-evoked the release of IL-1β、IL-6 and TNF-α were also partly inhibited by MRS2395 and MRS2211. Furthermore, ADPβS-evoked the increased expression of Iba-1, IL-1β、IL-6 and TNF-α mRNA and the release of IL-1β、IL-6 and TNF-α were nearly all blocked after co-administration of MRS2395 and MRS2211. Conclusion： P2Y12/P2Y13 receptor activation participates in the ADPβs- evoked the increased production of Iba-1, IL-1β、IL-6 and TNF-αin cultured spinal microglia cells.