Ethanol stimulates ciliary beating by dual cyclic nucleotide kinase activation in bovine bronchial epithelial cells
Ethanol stimulates ciliary beating by dual cyclic nucleotide kinase activation in bovine bronchial epithelial cells
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DOI：
10.1016/s0002-9440(10)63475-x
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发表时间：
2003-09-01
影响因子：
6
通讯作者：
Sisson, JH
中科院分区：
文献类型：
--
作者：
Wyatt, TA;Forgèt, MA;Sisson, JH
先前，我们已经证明乙醇（ROH）通过激活PKA刺激牛支气管上皮细胞（BBECs）的纤毛搏动频率（CBF）的快速增加。我们还发现，一氧化氮合酶抑制剂可阻断乙氧乙酸刺激的CBF增加。我们假设EtOH通过激活PKA和PKG途径急性刺激CBF。利用化学发光法检测一氧化氮（NO），我们直接测量了100 mmol/L EtOH处理BBECs 25分钟后NO产量的增加情况。用guanyyl环化酶抑制剂ODQ或LY83583预处理BBECs可抑制etoh刺激的CBF。低浓度（1 nmol/L）的环核苷酸类似物不会刺激CBF增加。然而，1 nmol/L的8Br-cAMP和8Br-cGMP的组合刺激了基线CBF的显著增加。交叉激活PKA和PKG需要非常高浓度的8Br-cAMP或8Br-cGMP（大于或等于100 μ mol/L），这表明在能够刺激CBF的浓度（1 nmol/L）下，cGMP不会发生PKA的交叉激活。8- pcpt -cGMP是cGMP的拮抗剂类似物，可阻断etoh刺激的PKA活性增加。这些数据支持etoh刺激的CBF增加需要PKA（通过cAMP）和PKG（通过NO）的双重激活。
Previously, we have shown that ethanol (ROH) stimulates a rapid increase in the ciliary beat frequency (CBF) of bovine bronchial epithelial cells (BBECs) via the activation of PKA. We have also shown that inhibitors of nitric oxide synthase block EtOH-stimulated increases in CBF. We hypothesize that EtOH acutely stimulates CBF via the activation of both PKA and PKG pathways. Using chemiluminescence detection of nitric oxide (NO), we directly measured increases in NO production in BBECs treated with 100 mmol/L of EtOH beginning at 25 minutes. Pretreatment of BBECs with guanylyl cyclase inhibitors, ODQ or LY83583, resulted in the inhibition of EtOH-stimulated CBF. Low concentrations (1 nmol/L) of cyclic nucleotide analogues do not stimulate CBF increases. However, a combination of both I nmol/L of 8Br-cAMP and 8Br-cGMP stimulates a significant increase over baseline CBF. This effect could be blocked by pretreating BBECs with inhibitors of either PKA or PKG. Very high concentrations of either 8Br-cAMP or 8Br-cGMP (greater than or equal to100 mumol/L) were required to cross-activate both PKA and PKG. This suggests that cross-activation of PKA by cGMP is not occurring at the concentrations (1 nmol/L) capable of stimulating CBF. 8-pCPT-cGMPS, an antagonist analogue to cGMP, blocked EtOH-stimulated PKA activity increases. These data support that EtOH-stimulated increases in CBF require the dual activation of both PKA (via cAMP) and PKG (via NO).