Ethanol increases phosphodiesterase 4 activity in bovine bronchial epithelial cells
Ethanol increases phosphodiesterase 4 activity in bovine bronchial epithelial cells
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DOI：
10.1016/j.alcohol.2003.06.005
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发表时间：
2003-08-01
期刊：
影响因子：
2.3
通讯作者：
Wyatt, TA
中科院分区：
文献类型：
--
作者：
Forgèt, MA;Sisson, JH;Wyatt, TA
气道上皮细胞中的乙醇暴露增加环腺苷酸（cAMP）依赖性蛋白激酶（PKA）活性。PKA和环磷酸鸟苷（cGMP）依赖性蛋白激酶（PKG）的激活已被证明可以增加牛支气管上皮细胞（BBEC）的纤毛搏动频率（CBF）。我们已经表明，生物相关浓度的乙醇刺激增加CBF在一氧化氮依赖性的方式，通过升高cAMP水平和随后的PKA激活介导。这种乙醇驱动的CBF快速和短暂的增加发生在暴露于100 mM乙醇后15至30分钟。然而，在长时间暴露于100 MM乙醇（大于或等于6小时）后，CBF和PKA的催化活性恢复到基线水平。我们推测，环核苷酸依赖性磷酸二酯酶（PDE）的活性减弱乙醇刺激的纤毛运动的持续时间。通过直接测定催化活性来确定乙醇对BBECs中PDE活性的影响。当BBEC与100 mM乙醇孵育时，cAMP水平在1小时内显著增加，PKA活性相应增加。用100 mM乙醇处理BBEC可显著增加cAMP-PDE活性，4-It。3-异丁基-1-甲基黄嘌呤，Ro 20-1724和咯利普兰可抑制乙醇刺激的cAMP-PDE活性。这些药物抑制乙醇刺激的cAMP-PDE活性，并增加在相同条件下观察到的乙醇刺激的PKA活性的幅度。这些发现支持了急性暴露（
Ethanol exposure in airway epithelium increases cyclic AMP (cAMP)-dependent protein kinase (PKA) activity. Activation of PKA and cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG) has been shown to increase ciliary beat frequency (CBF) in bovine bronchial epithelial cells (BBECs). We have shown that biologically relevant concentrations of ethanol stimulate increases in CBF in a nitric oxide-dependent manner, mediated through elevated cAMP levels and subsequent PKA activation. This ethanol-driven rapid and transient increase in CBF occurs 15 to 30 min after exposure to 100 mM ethanol. However, after prolonged exposure to 100 MM ethanol (greater than or equal to6 h), CBF and the catalytic activity of PKA return to baseline levels. We hypothesize that cyclic nucleotide-dependent phosphodiesterase (PDE) activity attenuates the duration of ethanol-stimulated ciliary motility. The effect of ethanol on the PDE activity in BBECs was determined through direct assay of catalytic activity. When BBECs were incubated with 100 mM ethanol, significant increases in cAMP levels occurred within I h, with corresponding increases in PKA activity. Treatment of BBECs with 100 mM ethanol increased cAMP-PDE activity significantly by 4 It. 3-Isobutyl-1-methylxanthine, Ro 20-1724, and rolipram inhibited ethanol-stimulated cAMP-PDE activity. These agents inhibited ethanol-stimulated cAMP-PDE activity and increased the magnitude of ethanol-stimulated PKA activity observed under the same conditions. These findings support the idea that acute exposure (