Department of Cardiology, The Affiliated Hospital of Medical School of Ningbo University, 247 Renmin Road, Ningbo, 315000, China.
Department of Pharmacology, Ningbo University School of Medicine, 818 Fenghua Road, Ningbo, 315000, China.
J Cell Sci. 2022 Jul 1;135(13). doi: 10.1242/jcs.259544. Epub 2022 Jul 8.
Macrophage polarization plays a key role in the inflammatory response. Various ion channels expressed in macrophages have been documented, but very little is known about their roles in macrophage polarization. We found that knockdown or blockade of the Kir2.1 (also known as KCNJ2) channel significantly inhibited M1 macrophage polarization, but promoted M2 macrophage polarization. Lipopolysaccharide (LPS)-induced M1 polarization was also remarkably suppressed in high extracellular K+ solutions (70 mM K+), and this inhibition was partially abolished by adding Ca2+ to the culture medium. Ca2+ imaging showed that Ca2+ influx was dependent on the hyperpolarized membrane potential generated by the Kir2.1 channel. The upregulation of phospho (p)-CaMK II, p-ERK, and p-NF-κB proteins in macrophages from the RAW264.7 cell line that were stimulated with LPS was significantly reversed by blocking the Kir2.1 channel or culturing the cells with 70 mM K+ medium. Furthermore, in vivo studies showed that mice treated with a Kir2.1 channel blocker were protected from LPS-induced peritonitis. In summary, our data reveal the essential role of the Kir2.1 channel in regulating macrophage polarization via the Ca2+/CaMK II/ERK/NF-κB signaling pathway.
巨噬细胞极化在炎症反应中起着关键作用。已经记录了各种在巨噬细胞中表达的离子通道,但对于它们在巨噬细胞极化中的作用知之甚少。我们发现,Kir2.1(也称为 KCNJ2)通道的敲低或阻断显著抑制了 M1 巨噬细胞极化,但促进了 M2 巨噬细胞极化。高细胞外 K+溶液(70 mM K+)也显著抑制了脂多糖(LPS)诱导的 M1 极化,而在培养基中添加 Ca2+可以部分消除这种抑制。Ca2+成像显示 Ca2+内流依赖于 Kir2.1 通道产生的超极化膜电位。用 LPS 刺激 RAW264.7 细胞系中的巨噬细胞后,磷酸化(p)-CaMK II、p-ERK 和 p-NF-κB 蛋白的上调被阻断 Kir2.1 通道或用 70 mM K+培养基培养细胞显著逆转。此外,体内研究表明,用 Kir2.1 通道阻断剂治疗的小鼠可以防止 LPS 诱导的腹膜炎。总之,我们的数据揭示了 Kir2.1 通道通过 Ca2+/CaMK II/ERK/NF-κB 信号通路调节巨噬细胞极化的重要作用。