巨噬細胞的胞外陷阱(MET)是最近新發現的巨噬細胞的複雜防禦機制，有別於吞噬作用，涉及DNA和抗菌蛋白的釋放。 它在清除病原體方面發揮著重要作用，但仍存在許多未知之處。 我們的研究發現，在脂多醣(LPS)刺激下，巨噬細胞會活化鈣池調控鈣離子流(SOCE)，激活 Orai 通道以促進鈣流入。 細胞質鈣離子的升高促使NADPH氧化酶(NOX)產生超氧陰離子，最終導致MET的形成。 此外，我們使用digitonin作為Triton X-100在免疫螢光染色的替代品，並結合使用lamin B抗體進行免疫螢光染色，創新地鑑定了METosis細胞。Digitonin的有限穿透性能確保核內抗體僅在 MET 細胞中著色，從而提供簡單直觀的分化方法。 此外，多數文獻注重鈣離子在PAD4依賴性的MET形成中的作用，但忽略了其在NOX依賴性中的重要性。 然而，我們的研究表明，鈣離子調節在NOX依賴性的MET生產中也起著至關重要的作用，挑戰了現有的觀點，並為未來的研究提供了新的思維。

Macrophage extracellular traps (MET) represent recently discovered complex defense mechanisms distinct from phagocytosis, which involve the release of DNA and antibacterial proteins. It plays an important role in removing pathogens, but many unknowns remain. Our investigation found that under lipopolysaccharide (LPS) stimulation, the macrophages undergo store-operated calcium entry (SOCE), activating Orai channels to facilitate calcium influx. The elevation of cytoplasmic calcium ions prompts the generation of superoxide anions by NADPH oxidase (NOX), ultimately leading to the formation of MET. Furthermore, we have innovatively identified METosis cells using digitonin as an alternative to triton X-100, coupled with immunofluorescent staining employing lamin antibodies. The limited permeability of digitonin ensures intranuclear antibody coloration exclusively in MET cells, providing a straightforward and intuitive differentiation method. In summary, prevailing literature acknowledges the role of calcium in PAD4-dependent MET formation while downplaying its significance in NOX-dependence. Our research, however, unveils that calcium regulation of NOX also plays a crucial role in MET production, challenging the existing perspectives and providing new ideas for future research.

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