Mass Spectrometry Resource, Division of Endocrinology, Metabolism, and Lipid Research, Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Biol Chem. 2012 Feb 17;287(8):5528-41. doi: 10.1074/jbc.M111.285114. Epub 2011 Dec 22.
Group VIA phospholipase A(2) (iPLA(2)β) in pancreatic islet β-cells participates in glucose-stimulated insulin secretion and sarco(endo)plasmic reticulum ATPase (SERCA) inhibitor-induced apoptosis, and both are attenuated by pharmacologic or genetic reductions in iPLA(2)β activity and amplified by iPLA(2)β overexpression. While exploring signaling events that occur downstream of iPLA(2)β activation, we found that p38 MAPK is activated by phosphorylation in INS-1 insulinoma cells and mouse pancreatic islets, that this increases with iPLA(2)β expression level, and that it is stimulated by the iPLA(2)β reaction product arachidonic acid. The insulin secretagogue D-glucose also stimulates β-cell p38 MAPK phosphorylation, and this is prevented by the iPLA(2)β inhibitor bromoenol lactone. Insulin secretion induced by d-glucose and forskolin is amplified by overexpressing iPLA(2)β in INS-1 cells and in mouse islets, and the p38 MAPK inhibitor PD169316 prevents both responses. The SERCA inhibitor thapsigargin also stimulates phosphorylation of both β-cell MAPK kinase isoforms and p38 MAPK, and bromoenol lactone prevents both events. Others have reported that iPLA(2)β products activate Rho family G-proteins that promote MAPK kinase activation via a mechanism inhibited by Clostridium difficile toxin B, which we find to inhibit thapsigargin-induced β-cell p38 MAPK phosphorylation. Thapsigargin-induced β-cell apoptosis and ceramide generation are also prevented by the p38 MAPK inhibitor PD169316. These observations indicate that p38 MAPK is activated downstream of iPLA(2)β in β-cells incubated with insulin secretagogues or thapsigargin, that this requires prior iPLA(2)β activation, and that p38 MAPK is involved in the β-cell functional responses of insulin secretion and apoptosis in which iPLA(2)β participates.
胰岛β细胞中的 VIA 组磷酯酶 A2(iPLA2β)参与葡萄糖刺激的胰岛素分泌和肌浆内质网 ATP 酶(SERCA)抑制剂诱导的细胞凋亡,而这两种作用都可被 iPLA2β 活性的药理学或遗传学降低所减弱,也可被 iPLA2β 的过度表达所增强。在研究 iPLA2β 激活后发生的信号事件时,我们发现 p38MAPK 在 INS-1 胰岛细胞瘤和小鼠胰岛中通过磷酸化而被激活,这种激活作用随 iPLA2β 表达水平的增加而增强,并受 iPLA2β 反应产物花生四烯酸的刺激。胰岛素分泌激动剂 D-葡萄糖也刺激β细胞 p38MAPK 磷酸化,而这种磷酸化可被 iPLA2β 抑制剂溴烯醇内酯所阻止。在 INS-1 细胞和小鼠胰岛中,过表达 iPLA2β 可放大 D-葡萄糖和佛波醇 12,13-二丁酸酯诱导的胰岛素分泌,而 p38MAPK 抑制剂 PD169316 可阻止这两种反应。SERCA 抑制剂 thapsigargin 也可刺激两种β细胞 MAPK 激酶同工型和 p38MAPK 的磷酸化,而溴烯醇内酯可阻止这两种反应。其他人已经报道 iPLA2β 产物可激活 Rho 家族 G 蛋白,通过一种被艰难梭菌毒素 B 抑制的机制促进 MAPK 激酶的激活,我们发现艰难梭菌毒素 B 可抑制 thapsigargin 诱导的β细胞 p38MAPK 磷酸化。p38MAPK 抑制剂 PD169316 还可阻止 thapsigargin 诱导的β细胞凋亡和神经酰胺的生成。这些观察结果表明,在与胰岛素分泌激动剂或 thapsigargin 孵育的β细胞中,p38MAPK 在 iPLA2β 之后被激活,这需要 iPLA2β 的预先激活,并且 p38MAPK 参与 iPLA2β 参与的胰岛素分泌和细胞凋亡的β细胞功能反应。