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探讨Elmo1在中性粒细胞胞外诱捕网(neutrophil extracellular trap,NET)形成过程中的作用机制。分别提取WT小鼠和Elmo1敲除小鼠的中性粒细胞后,用PMA诱导NET形成。运用激光扫描共聚焦显微镜对NET的形成进行观察,用多功能酶标仪读取荧光强度值并进行定量分析。为了进一步探索Elmo1调节NET形成的分子机制,我们用Western blotting检测了NET形成过程中相关信号通路的激活情况,包括p38和Akt。结果发现,Elmo1敲除的条件下,NET的形成明显减少。同时,在Elmo1敲除小鼠的中性粒细胞中,PMA诱发的p38和Akt的激活也显著降低。抑制p38、Akt后,WT和Elmo1敲除型小鼠之间NET形成的差异消失。说明Elmo1通过p38和Akt参与上调NET的形成。上述结果提示Elmo1参与上调NET形成。
To investigate the mechanism of Elmo1 in neutrophil extracellular trap (NET) formation. After extracting neutrophils from WT mice and Elmo1 knockout mice respectively, NET formation was induced with PMA. The formation of NET was observed by laser scanning confocal microscopy. The fluorescence intensity was read by multifunctional microplate reader and quantified. To further explore the molecular mechanism by which Elmo1 regulates NET formation, we examined the activation of related signaling pathways during NET formation by Western blotting, including p38 and Akt. The results showed that, Elmo1 knockout conditions, NET significantly reduced. In the meantime, PMA-induced activation of p38 and Akt was also significantly reduced in Elmo 1 knockout mouse neutrophils. After inhibition of p38, Akt, differences in NET formation between WT and Elmo1 knockout mice disappeared. That Elmo1 through p38 and Akt involved in the up-regulation of NET formation. These results suggest that Elmo1 is involved in the up-regulation of NET formation.