THP-1细胞中p2x7R对ISG20的调节作用

发布时间：2018-06-11 17:55

本文选题：嘌呤能受体 + 白介素β　；参考：《免疫学杂志》2017年09期

【摘要】：目的探讨脂多糖诱导的THP-1源性巨噬细胞中,嘌呤能受体2x7(p2x7R)对IFN-α诱导干扰素刺激基因20(ISG20)的影响及其可能的机制。方法 THP-1源性巨噬细胞经LPS诱导4 h后分别加入p2x7R非特异性抑制剂苏拉明(Suramin)、磷酸吡哆醛(PPADS),p2x7R特异性抑制剂oxidized ATP(O-ATP)、亮蓝G(BBG)、以及p2x7R特异性激动剂苯甲酰苯甲酸ATP(BzATP)作用1 h后,联合IFN-α培养24 h。监测细胞内ISG20 mRNA、蛋白的表达差异及上清液中IL-1β的含量。BzATP组中,观察不同浓度BzATP以及caspase-1抑制剂预处理1 h后Bz ATP、IFN-α对ISG20的调节作用。结果 RT-PCR及Western blot结果显示,p2x受体广谱抑制剂与p2x7R的特异性抑制剂具有抑制IFN-α对ISG20的诱导作用,IFN-α联用抑制剂组与IFN-α联用激动剂BzATP组中ISG20 mRNA较IFN-α组差异有统计学意义(P0.05)。浓度梯度实验中,上清液中IL-1β的含量在加入BzATP、BBG后发生改变。另外,Bz ATP直接诱导的ISG20可被caspase-1抑制剂抑制,上清液中IL-1β同样受到抑制。结论在THP-1细胞中,BzATP对ISG20的诱导作用不同于IFN-α途径直接诱导ISG20的生成。这为将来探索新的抗病毒治疗提供了新的思路。
[Abstract]:Objective to investigate the effect of purinergic receptor 2x7 (p2x7R) on IFN- 伪 -induced interferon stimulating gene 20ISG20 in THP-1 derived macrophages induced by lipopolysaccharide (LPS) and its possible mechanism. Methods THP-1 derived macrophages were treated with p2x7R nonspecific inhibitor Suraminine, p2x7R specific inhibitor oxidized ATP O-ATPase, brilliant blue GBS GBS, and p2x7R specific agonist ATP BzATP for 1 h. Combined with IFN- 伪 for 24 h. To monitor the difference of ISG20 mRNAs and protein expression and the content of IL-1 尾 in supernatant. In BzATP group, we observed the regulation of ISG20 by Bz ATP and caspase-1 inhibitor pretreatment for 1 h. Results the results of RT-PCR and Western blot showed that P2x receptor broad-spectrum inhibitor and p2x7R specific inhibitor could inhibit the induction of ISG20 by IFN- 伪 and IFN- 伪 combined with IFN- 伪 agonist BzATP. There was a significant difference in ISG20 mRNA between IFN- 伪 group and IFN- 伪 group compared with IFN- 伪 group. In the concentration gradient experiment, the content of IL-1 尾 in the supernatant changed after adding BzATP BBG. In addition, ISG20 directly induced by BZ ATP could be inhibited by caspase-1 inhibitor, and IL-1 尾 in supernatant was also inhibited. Conclusion the induction of ISG20 by BzATP in THP-1 cells is different from that induced by IFN- 伪 pathway. This provides a new idea for exploring new antiviral therapy in the future.
【作者单位】：重庆医科大学附属第一医院感染科;
【基金】：国家自然科学基金(81271838)
【分类号】：R392

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