Thrombin induces a temporal biphasic vascular response through the differential phosphorylation of endothelial nitric oxide synthase via protease-activated receptor-1 and protein kinase C
Thrombin induces a temporal biphasic vascular response through the differential phosphorylation of endothelial nitric oxide synthase via protease-activated receptor-1 and protein kinase C
复制标题
凝血酶通过蛋白酶激活受体 1 和蛋白激酶 C 内皮一氧化氮合酶的差异磷酸化诱导暂时性双相血管反应
DOI：
10.1016/j.jphs.2022.02.001
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发表时间：
2022
影响因子：
3.5
通讯作者：
et al
中科院分区：
文献类型：
--
作者：
Akihiro Okamura;Hisatome I;et al
内皮一氧化氮合酶 (eNOS) 是一种关键的调节酶，通过产生一氧化氮来控制血管张力。尽管凝血酶也主要通过激活蛋白酶激活受体 (PAR) 来调节血管张力，但凝血酶如何控制 eNOS 酶活性的时间过程和机制尚不清楚。 eNOS 酶活性因 eNOS-Ser1177 磷酸化而增强，并因 eNOS-Thr495 磷酸化而降低。在这项研究中，我们假设凝血酶通过 eNOS 的差异磷酸化来调节血管张力。使用大鼠降主动脉，我们发现凝血酶通过激活的 PAR-1 以 eNOS 依赖性方式调节血管张力。我们还表明凝血酶会引起暂时的双相反应。蛋白激酶 C (PKC) 与凝血酶诱导反应的第二阶段相关。蛋白质印迹分析表明，人脐静脉内皮细胞中凝血酶磷酸化 eNOS-Ser1177 和 eNOS-Thr495。 PKC 抑制剂可抑制凝血酶诱导的 eNOS-Thr495 磷酸化，但不能抑制 eNOS-Ser1177 磷酸化。我们的结果表明，凝血酶通过活化的 PAR-1 对 eNOS 进行差异性磷酸化，从而诱导暂时性双相血管反应。凝血酶通过 eNOS-Ser1177 的磷酸化引起短暂的血管舒张，随后通过 PKC 通过 eNOS-Thr495 的磷酸化来减弱血管舒张，从而调节血管张力。
Endothelial nitric oxide synthase (eNOS) is a critical regulatory enzyme that controls vascular tone via the production of nitric oxide. Although thrombin also modulates vascular tone predominantly via the activation of protease-activated receptors (PARs), the time course and mechanisms involved in how thrombin controls eNOS enzymatic activity are unknown. eNOS enzymatic activity is enhanced by the phosphorylation of eNOS-Ser1177 and reduced by the phosphorylation of eNOS-Thr495. In this study, we hypothesized that thrombin regulates vascular tone through the differential phosphorylation of eNOS. Using rat descending aorta, we show that thrombin modulates vascular tone in an eNOS-dependent manner via activated PAR-1. We also show that thrombin causes a temporal biphasic response. Protein kinase C (PKC) is associated with second phase of thrombin-induced response. Western blot analysis demonstrated thrombin phosphorylated eNOS-Ser1177 and eNOS-Thr495 in human umbilical vein endothelial cells. A PKC inhibitor suppressed the thrombin-induced phosphorylation of eNOS-Thr495, but not that of eNOS-Ser1177. Our results suggest that thrombin induces a temporal biphasic vascular response through the differential phosphorylation of eNOS via activated PAR-1. Thrombin causes transient vasorelaxation by the phosphorylation of eNOS-Ser1177, and subsequent attenuation of vasorelaxation by the phosphorylation of eNOS-Thr495 via PKC, leading to the modulation of vascular tone.