间质巨噬细胞 (IM) 对于肺组织中的器官稳态、炎症和自主免疫反应至关重要，这是通过极化为促炎 M1 和 M2 状态以进行组织修复来实现的。然而，它们的远程实质定位和低计数是其分离和组织炎症期间特定作用的分子表征的限制因素。我们通过将小鼠与基质细胞共培养来分离出足够数量的活鼠IM，并分析了应用脂多糖（LPS）和干扰素γ后初始和M1极化IM中瞬时受体电位（TRP）通道的mRNA表达模式。 TRP 通道褪黑素家族第二个成员 TRPM2 的 M-RNA 在 M1 状态下上调，并且通过其特异性激活剂 ADP-核糖诱导的特征电流来识别功能通道。最有趣的是，与 WT 细胞相比，M1 极化但 TRPM2 缺陷的 IM 中细胞因子的产生和白介素 1α (IL-1α)、IL-6 和肿瘤坏死因子 α 的分泌显着增强。 ROS 产生的 H2O2 从线粒体释放的 ADP-核糖 (ADPR) 激活 TRPM2 通道，显着增加质膜去极化，从而抑制 NADPH 氧化酶产生活性氧，并减少负反馈环中细胞因子的产生和分泌。因此，TRPM2 通道对于 M1 极化小鼠 IM 中细胞因子产生的调节至关重要。这些通道的特异性激活可能会促进抗炎表型，并防止在 COVID-19 患者中经常观察到的有害细胞因子风暴。© 2024 作者。 《细胞生理学杂志》由 Wiley periodicals LLC 出版。

Interstitial macrophages (IMs) are essential for organ homeostasis, inflammation, and autonomous immune response in lung tissues, which are achieved through polarization to a pro-inflammatory M1 and an M2 state for tissue repair. Their remote parenchymal localization and low counts, however, are limiting factors for their isolation and molecular characterization of their specific role during tissue inflammation. We isolated viable murine IMs in sufficient quantities by coculturing them with stromal cells and analyzed mRNA expression patterns of transient receptor potential (TRP) channels in naïve and M1 polarized IMs after application of lipopolysaccharide (LPS) and interferon γ. M-RNAs for the second member of the melastatin family of TRP channels, TRPM2, were upregulated in the M1 state and functional channels were identified by their characteristic currents induced by ADP-ribose, its specific activator. Most interestingly, cytokine production and secretion of interleukin-1α (IL-1α), IL-6 and tumor necrosis factor-α in M1 polarized but TRPM2-deficient IMs was significantly enhanced compared to WT cells. Activation of TRPM2 channels by ADP-ribose (ADPR) released from mitochondria by ROS-produced H2O2 significantly increases plasma membrane depolarization, which inhibits production of reactive oxygen species by NADPH oxidases and reduces cytokine production and secretion in a negative feedback loop. Therefore, TRPM2 channels are essential for the regulation of cytokine production in M1-polarized murine IMs. Specific activation of these channels may promote an anti-inflammatory phenotype and prevent a harmful cytokine storm often observed in COVID-19 patients.© 2024 The Author(s). Journal of Cellular Physiology published by Wiley Periodicals LLC.