P2X7 receptor (P2X7R) is an ATP-gated non-selective cation channel which mediates ATP-induced inflammation in macrophages. Transient receptor potential (TRP) receptors are nociceptors in cellular membrane which can perceive the stimuli of environmental irritant. The interaction between TRP channels and P2X7R has been found while the details about inflammation are still unclear. In this study, we suggested that transient receptor potential ankyrin 1 (TRPA1), a member of TRP superfamily, participates in ATP-induced oxidative stress and inflammation in human acute monocytic leukemia cell line (THP-1)-derived macrophage. The co-localization between TRPA1 and P2X7R was detected using immunofluorescence in THP-1-derived macrophage and transfected human embryonic kidney cell line (HEK293T). The mechanism by which ATP or 3′-O-(4-Benzoylbenzoyl)-ATP (BzATP) induces the activation of macrophages was verified by calcium imaging, mitochondrial reactive oxygen species (mtROS) detection, mitochondrial membrane potential (∆Ψ_m) measurement, flow cytometry, enzyme-linked immunosorbent assay (ELISA), western blotting, CCK-8 assay, and the lactate dehydrogenase (LDH) release cytotoxic assay. The BzATP and ATP induced calcium overload, mitochondria injury, interleukin-1β (IL-1β) secretion, and cytotoxicity can be inhibited by TRPA1 antagonists. These results indicated that TRPA1 can co-localize with P2X7R and mediate ATP-induced oxidative stress and inflammation. Therefore, the inhibition of TRPA1 may provide a potential therapy for ATP-elicited inflammatory diseases, including atherosclerosis.

P2X7受体（P2X7R）是ATP门控的非选择性阳离子通道，在巨噬细胞中介导ATP诱导的炎症。瞬态受体电位（TRP）受体是细胞膜中的伤害感受器，可以感知环境刺激物。已经发现TRP通道和P2X7R之间的相互作用，但有关炎症的细节仍不清楚。在这项研究中，我们建议，瞬时受体电位锚蛋白1（TRPA1），TRP超家族的成员，参与人急性单核细胞白血病细胞系（THP-1）衍生的巨噬细胞中ATP诱导的氧化应激和炎症。在THP-1衍生的巨噬细胞和转染的人胚胎肾细胞系（HEK293T）中使用免疫荧光检测了TRPA1和P2X7R之间的共定位。ATP或3'- O的机制-（4-苄基苯甲酰基）-ATP（BzATP）诱导巨噬细胞活化已通过钙成像，线粒体活性氧（mtROS）检测，线粒体膜电位（∆Ψ _m）测量，流式细胞仪，酶联免疫吸附测定（ ELISA），蛋白质印迹法，CCK-8测定法和乳酸脱氢酶（LDH）释放细胞毒性测定法。BzATP和ATP诱导的钙超载，线粒体损伤，白介素1β（IL-1β）分泌和细胞毒性可被TRPA1拮抗剂抑制。这些结果表明TRPA1可以与P2X7R共定位并介导ATP诱导的氧化应激和炎症。因此，抑制TRPA1可能为ATP引起的炎性疾病（包括动脉粥样硬化）提供潜在的治疗方法。