Estradiol-mediated endothelial nitric oxide synthase association with heat shock protein 90 requires adenosine monophosphate-dependent protein kinase
Estradiol-mediated endothelial nitric oxide synthase association with heat shock protein 90 requires adenosine monophosphate-dependent protein kinase
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DOI：
10.1161/circulationaha.105.546812
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发表时间：
2005-06-28
期刊：
影响因子：
37.8
通讯作者：
Keaney, JF
中科院分区：
文献类型：
--
作者：
Schulz, E;Anter, E;Keaney, JF
背景-雌激素通过涉及雌激素受体-α（ER α）、蛋白激酶B/ Akt、促分裂原活化蛋白激酶和热休克蛋白90（HSP 90）的机制激活内皮型一氧化氮合酶（eNOS）。最近，AMP活化的蛋白激酶（AMPK），一种在细胞适应代谢应激中起关键作用的酶，已经被牵连在由激素脂联素和胰岛素引起的生理性eNOS活化中。因此，我们研究了AMPK是否被内皮细胞中的雌二醇激活，并在雌二醇诱导的eNOS激活中发挥作用.方法和结果--通过AMPK及其下游靶乙酰辅酶A羧化酶对雌二醇的反应（1 nmol/ L至10 μ mol/ L，1至30分钟）的磷酸化测定，猪主动脉内皮细胞表现出时间和浓度依赖性AMPK激活.雌二醇激活AMPK不依赖于AMP水平和ER α，但需要雌二醇转化为其儿茶酚代谢物。内皮素处理增加了eNOS催化活性，当内皮细胞被AMPK显性阴性腺病毒感染时，这种作用在很大程度上被逆转。然而，AMPK的抑制并不改变雌二醇诱导的eNOS在丝氨酸1177或苏氨酸495处的磷酸化，但降低了eNOS与HSP 90的相互作用。与这一观察结果相一致的是，来自α（1）- AMPK缺失小鼠的血管显示出对雌二醇应答的eNOS介导的NO产生缺陷.结论--综合起来，这些数据表明AMPK活性对于雌二醇诱导的eNOS活化是必需的，通过促进eNOS与HSP 90的相互作用.这些数据表明AMPK在调节内皮细胞NO生物活性和HSP 90功能中的新作用。
Background - Estradiol activates endothelial nitric oxide synthase ( eNOS) by mechanisms that involve estrogen receptor-alpha ( ER alpha), protein kinase B/ Akt, mitogen- activated protein kinases, and heat shock protein 90 ( HSP90). Recently, AMP- activated protein kinase ( AMPK), an enzyme that plays a crucial role in cellular adaptation to metabolic stress, has been implicated in physiological eNOS activation by the hormones adiponectin and insulin. We therefore investigated whether AMPK is activated by estradiol in endothelial cells and plays a role in estradiol- induced eNOS activation.Methods and Results - Porcine aortic endothelial cells exhibited time- and concentration- dependent AMPK activation as determined by phosphorylation of AMPK and its downstream target acetyl coenzyme A carboxylase in response to estradiol ( 1 nmol/ L to 10 mu mol/ L, 1 to 30 minutes). AMPK activation by estradiol was independent of both AMP levels and ER alpha but required estradiol conversion to its catechol metabolites. Estradiol treatment increased eNOS catalytic activity, an effect that was largely reversed when endothelial cells were infected with an AMPK dominant- negative adenovirus. However, inhibition of AMPK did not alter estradiol- induced eNOS phosphorylation at serine 1177 or threonine 495 but decreased eNOS interaction with HSP90. Consistent with this observation, blood vessels from alpha(1)- AMPK - null mice exhibited defective eNOS- mediated NO production in response to estradiol.Conclusions - Taken together, these data indicate that AMPK activity is essential for estradiol- induced eNOS activation via the promotion of eNOS interaction with HSP90. These data point to a novel role for AMPK in modulating endothelial cell NO bioactivity and HSP90 function.