ANO1通过MEK/ERK/ELK1途径介导百草枯诱导巨噬细胞NLRP3炎性小体活化作用机制

NLRP3炎性小体介导百草枯致巨噬细胞活化机制不清。我们研究发现百草枯可以激活巨噬细胞钙激活氯通道ANO1和NLRP3炎性小体以及MEK/ERK/ELK1信号通路；使用ANO1抑制剂可以抑制MEK/ERK/ELK1信号通路及NLRP3炎性小体活化。通过生物学软件预测在NLRP3启动子区域存在着ELK1的结合位点。因此，我们推测ANO1可能通过MEK/ERK/ELK1途径介导百草枯诱导巨噬细胞NLRP3炎性小体活化。本研究将采用基因过表达和沉默技术，明确ANO1对百草枯诱导的巨噬细胞NLRP3炎性小体活化调控作用；采用MEK/ERK/ELK1通路抑制剂明确该通路在此过程中起到的关键作用；采用双荧光素酶报告基因系统，确定ELK1对NLRP3基因的启动作用以及关键结合位点。最终阐明ANO1调控百草枯诱导巨噬细胞NLRP3炎性小体活化的作用机制，为临床上百草枯中毒的治疗提供新靶点。

The mechanisms of NLRP3 inflammasome-mediated inflammation in macrophage induced by paraquat is unclear. We found that the calcium-activated chloride channel ANO1 and NLRP3 inflammasome as well as MEK/ERK/ELK1 pathway were activated in paraquat-induced macrophages; the activation of MEK/ERK/ELK1 pathway and NLRP3 inflammasome was reduced by the ANO1 inhibitor. It was predicted that there were the ELK1 binding sites in the promoter region of NLRP3 using the biological software. Therefore, we hypothesize that ANO1 might regulate NLRP3 inflammasome activation in paraquat-induced macrophage via MEK / ERK / ELK1 pathway. In this study, we will use gene overexpression/silence technology to clarify the mechanism of ANO1 regulated NLRP3 inflammasome activation in paraquat-induced macrophages. Additionally, we will use the inhibitors of MEK / ERK / ELK1 pathway illustrate its key role in the regulation of NLRP3 inflammasome activation. Finally, we will use the dual-luciferase report gene system to determine the promotor of NLRP3 gene and its key binding site of ELK1. In the end, the mechanisms that ANO1regulated NLRP3 inflammasome activation in paraquat-induced macrophage would be illustrated, and the valuable new targets for paraquat poisoning therapy will be provided.