Department of Physiology and Pharmacology, University of Toledo College of Medicine, Health Science Campus, 3000 Arlington Av., Toledo, OH 43614, USA.
Biochem Biophys Res Commun. 2012 Oct 12;427(1):178-84. doi: 10.1016/j.bbrc.2012.09.038. Epub 2012 Sep 16.
We previously showed that survival signaling in TNFα-treated, human THP1-derived macrophages (TDMs) has an obligatory requirement for constitutive Ca(2+) influx through a mechanism involving calmodulin/calmodulin kinase II (CAM/CAMKII). We also demonstrated that such requirement also applies to the protective actions of TNFα in murine bone marrow-derived macrophages (BMDMs) and that TRPC3 channels mediate constitutive Ca(2+) influx. Using a pharmacological approach we here examined if in BMDMs, similarly to TDMs, TNFα-induced survival signaling also involves CAM/CAMKII. In BMDMs, TNFα induced rapid activation of the survival pathways NFκB, AKT and p38MAPK. All these routes were activated in a PI3K-dependent fashion. Activation of AKT and NFκB, but not that of p38MAPK, was abrogated by the CAM inhibitor W7, while KN-62, a CAMKII inhibitor, prevented activation of AKT and p38MAPK but not that of NFκB. Inhibition of CAM or CAMKII completely prevented the protective actions of TNFα. Our observations indicate that in BMDMs CAM and CAMKII have differential contributions to the components of TNFα-dependent survival signaling and underscore a complex interplay among canonical survival routes. These findings set a signaling framework to understand how constitutive Ca(2+) influx couples to macrophage survival in BMDMs.
我们之前的研究表明,在 TNFα 处理的人 THP1 衍生巨噬细胞(TDMs)中,生存信号需要通过一种涉及钙调蛋白/钙调蛋白激酶 II(CAM/CAMKII)的机制来强制进行持续的 Ca2+内流。我们还证明,这种要求也适用于 TNFα 在小鼠骨髓来源的巨噬细胞(BMDMs)中的保护作用,并且 TRPC3 通道介导持续的 Ca2+内流。在此,我们使用药理学方法研究了在 BMDMs 中,TNFα 诱导的生存信号是否也涉及 CAM/CAMKII。在 BMDMs 中,TNFα 诱导了 NFκB、AKT 和 p38MAPK 等生存途径的快速激活。所有这些途径均以 PI3K 依赖性方式激活。CAM 抑制剂 W7 阻断 AKT 和 NFκB 的激活,但不阻断 p38MAPK 的激活,而 CAMKII 抑制剂 KN-62 则阻止 AKT 和 p38MAPK 的激活,但不阻止 NFκB 的激活。CAM 或 CAMKII 的抑制完全阻止了 TNFα 的保护作用。我们的观察结果表明,在 BMDMs 中,CAM 和 CAMKII 对 TNFα 依赖性生存信号的组成部分有不同的贡献,并强调了经典生存途径之间的复杂相互作用。这些发现为理解在 BMDMs 中持续的 Ca2+内流如何与巨噬细胞的生存相关提供了一个信号框架。