LPS与ATP共同诱导巨噬细胞中NLRP3炎症小体的激活

Mechanisms of lipopolysaccharide induced the activation of NLRP3 inflammation immune response in macrophages

【目的】以脂多糖(LPS)为刺激源,探索小鼠巨噬细胞(RAW264.7)中NOD样受体家族pyrin结构域蛋白3(NLRP3)炎性小体激活的响应机制。【方法】试验分为对照组、LPS组、LPS与ATP共同刺激组。ELISA检测NLRP3源性白介素-1β(IL-1β)表达情况;RT-PCR检测NLRP3、Caspase-1、白介素-1β(IL-1β)的转录水平;ELISA检测LPS与NLRP3其他激动剂(MSU、CPPD、SiO2、Alum)共同作用后IL-1β的表达以及PI染色检测细胞焦亡。【结果】与对照组相比,LPS刺激组对IL-1β的表达无显著影响,LPS/ATP共同刺激组IL-1β的表达显著升高;LPS/ATP刺激组NLRP3、Caspase-1、IL-1βmRNA的表达显著升高且LPS/ATP刺激组发生明显的细胞焦亡。【结论】LPS与ATP共同作用能够显著提高NLRP3、Caspase-1、IL-1β的表达,说明LPS对NLRP3炎性小体的激活是LPS与ATP共同作用实现的。

【Objective】To explore the mechanisms of NLRP3inflammation immune response in macrophages,lipopolysaccharide(LPS)was used as stress source.【Methods】Three group of experiments were designed:control,LPS and LPS/ATP,respectively.The NLRP3-inflammation-derived IL-1βsecretion was measured by ELISA.The transcriptional level of NLRP3,Caspase-1,IL-1βwas measured by RT-PCR.We further examined whether IL-1βwas induced by other NLRP3inflammation activators,including monosodium urate(MSU),calcium pyrophosphate dihydrate(CPPD),SiO2nanoparticles,and alum crystals.Pyroptosis was measured by propidium iodide(PI).【Result】Compared with control group,the expression of IL-1βwas not obviously influenced by LPS group.The expression of IL-1β,as well as NLRP3and Caspase-1,in raw264.7macrophages was obviously induced by the combing of LPS and ATP;furthermore,pyroptosis was increased.【Conclusion】LPS and ATP induce the expression of NLRP3,Caspase-1,IL-1β.