Positive signaling interactions between arsenic and ethanol for angiogenic gene induction in human microvascular endothelial cells.
Positive signaling interactions between arsenic and ethanol for angiogenic gene induction in human microvascular endothelial cells.
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DOI：
10.1093/toxsci/kfn003
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发表时间：
2008-04
期刊：
影响因子：
--
通讯作者：
L. Klei;A. Barchowsky
中科院分区：
文献类型：
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作者：
L. Klei;A. Barchowsky
饮用水中的砷可能通过尚未解决的机制促进全世界数百万人的血管疾病。此外，人们对砷和其他常见的血管毒物（如酒精）共同接触的影响知之甚少。为了研究砷和醇之间的信号相互作用，在0.1%乙醇（EtOH）存在或不存在的情况下，将人微血管内皮（HMVEC）细胞暴露于无细胞毒性浓度的亚砷酸盐（1-5微米）中。共同暴露，而不是单独暴露于任何一种物质，迅速增加了Fyn酪氨酸激酶的活性，109 kda蛋白的酪氨酸磷酸化和蛋白激酶C (PKC) δ的丝氨酸磷酸化。109 kda蛋白被鉴定为PYK2，是血管整合素信号的调节因子和PKCdelta的上游激活因子。在15分钟内共暴露可增加磷脂酶Cgamma1的膜定位，但单独使用任何一种药物均不能增加。相比之下，两种药物均增加了Rac1-GTPase的膜定位。共同暴露，但不是单独暴露于任何一种药物，诱导血管生成基因，血管内皮细胞生长因子（Vegfa）和胰岛素样生长因子-1 （Igf1）的转录水平。然而，EtOH抑制砷诱导的核因子kappab驱动的白介素-8和胶原-1的表达。选择性PKC抑制剂对诱导基因表达的不同影响，加上缺乏诱导血红素加氧酶-1的相互作用，进一步证明了砷反应信号通路对EtOH相互作用的敏感性不同。最后，在体外血管生成实验中，共暴露增强了内皮管的形成。这些数据表明，砷和EtOH暴露之间发生复杂的相互作用，在功能上影响内皮信号，从而诱导基因和重塑刺激。
Arsenic in the drinking water may promote vascular diseases in millions of people worldwide through unresolved mechanisms. In addition, little is known of the effects of coexposures to arsenic and other common vasculature toxicants, such as alcohol. To investigate signaling interactions between arsenic and alcohols, primary human microvascular endothelial (HMVEC) cells were exposed to noncytotoxic concentrations of arsenite (1-5 microM) in the presence or absence of 0.1% ethanol (EtOH). Coexposure, but not exposure to either agent alone, rapidly increased active Fyn tyrosine kinase, tyrosine phosphorylation of a 109-kDa protein and serine phosphorylation of protein kinase C (PKC)delta. The 109-kDa protein was identified as PYK2, a regulator of vascular integrin signaling and an upstream activator of PKCdelta. Membrane localization of phospholipase Cgamma1 was increased by coexposure within 15 min, but not by either agent alone. In contrast, both agents equally increased membrane localization of Rac1-GTPase. Coexposure, but not exposure to either agent alone, induced transcript levels for the angiogenic genes, vascular endothelial cell growth factor (Vegfa) and insulin-like growth factor-1 (Igf1). However, EtOH inhibited arsenic-induced, nuclear factor-kappaB-driven interleukin-8 and collagen-1 expression. Differential effects of selective PKC inhibitors on induced gene expression combined with a lack of interaction for induction of hemeoxygenase-1 further demonstrated that arsenic-responsive signaling pathways differ in sensitivity to EtOH interactions. Finally, coexposure enhanced endothelial tube formation in in vitro angiogenesis assays. These data indicate that complex interactions occur between arsenic and EtOH exposures that functionally affect endothelial signaling for gene induction and remodeling stimuli.