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Activation of CaMKKβ/AMPKα pathway by 2‐AG in human platelets

机译:在人血小板中对2-AG的Camkkβ/AMPKα途径的激活

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Abstract The objective of this study was to determine whether AMPK is activated by 2‐arachidonoylglycerol (2‐AG) and participates to the cytoskeleton control in human platelets. We found that 2‐AG stimulates the AMPKα activation through a Ca 2+ /Calmodulin‐dependent pathway as the specific inhibition of the CaMKKβ by STO‐609 inhibits the AMPKα phosphorylation/activation. Moreover, the CaMKKβ/AMPKα pathway activated by 2‐AG is involved in the phosphorylation of cofilin, vasodilator stimulated phosphoprotein (VASP), and myosin light chain (MLCs). These proteins participate to actin cytoskeletal remodelling during aggregation. We found that the phosphorylation/activation inhibition of these proteins is associated with a significant reduction in actin polymerization, aggregation, ATP, and α‐granule secretion. Finally, AMPKα activation, Cofilin, VASP, and MLCs phosphorylation are significantly reduced by SR141716, the specific inhibitor of type 1 cannabinoid (CB1) receptor, suggesting that the CB1 receptor is involved in the 2‐AG effect. In conclusion, we have shown that the CaMKKβ/AMPKα pathway is activated by 2‐AG in human platelets and controls the phosphorylation of key proteins involved in actin polymerization and aggregation.
机译:摘要本研究的目的是确定AMPK是否由2-arachidonlgycerol(2-Ag)激活,并参与人血小板中的细胞骨架控制。我们发现,由于STO-609对CAMKKβ的特异性抑制抑制AMPKα磷酸化/活化,因此通过Ca 2+ /钙调蛋白依赖性途径刺激AMPKα激活刺激。此外,由2-AG激活的Camkkβ/AMPKα途径涉及Cofilin,血管扩张剂刺激磷蛋白(VASP)和肌蛋白轻链(MLC)的磷酸化。这些蛋白质在聚集期间参与肌骨骨骼重塑。我们发现这些蛋白质的磷酸化/活化抑制与肌动蛋白聚合,聚集,ATP和α-颗粒分泌的显着降低有关。最后,通过SR141716,SR141716,1型大麻素(CB1)受体的特异性抑制剂显着降低了AMPKα活化,CoFilin,VAS和MLC磷酸化,表明CB1受体参与2-AG效应。总之,我们已经表明,Camkkβ/Ampkα途径在人血小板中由2-Ag激活,并控制肌动蛋白聚合和聚集中涉及的关键蛋白质的磷酸化。

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