Adenosine activation of A2B receptor(s) is essential for stimulated epithelial ciliary motility and clearance
Adenosine activation of A2B receptor(s) is essential for stimulated epithelial ciliary motility and clearance
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DOI：
10.1152/ajplung.00203.2010
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发表时间：
2011-08-01
影响因子：
4.9
通讯作者：
Wyatt, Todd A.
中科院分区：
文献类型：
--
作者：
Allen-Gipson, Diane S.;Blackburn, Michael R.;Wyatt, Todd A.
Allen-Gipson DS，Blackburn MR，Schneider DJ，Zhang H，Bluitt DL，Jobell JC，Yanov D，Sisson JH，Wyatt TA。腺苷激活A（2B）受体对于刺激上皮纤毛运动和清除是必需的。美国生理学杂志肺细胞分子生理学301：L171-L180，2011年。首次发表于2011年5月27日; doi：10.1152/ajplung.00203.2010.-对于肺清除至关重要的粘液纤毛清除依赖于纤毛搏动频率（CBF）、纤毛的协调和纤毛周液的维持。腺苷是ATP的代谢产物，是纤毛运动的重要调节剂。然而，特异性腺苷受体对关键气道纤毛运动过程的贡献尚不清楚。我们假设腺苷通过激活其细胞表面受体（A（1）、A（2A）、A（2B）或A（3））来调节纤毛运动。为了验证这一假设，用已知的纤毛激活剂异丙肾上腺素刺激从野生型和腺苷受体敲除小鼠（分别为A（1）、A（2A）、A（2B）或A（3））切除的小鼠气管环（MTR）和牛纤毛支气管上皮细胞（BBEC（ISO; 10 μ M）和/或丙卡特罗（10 μ M），在存在或不存在5 ′-测定非选择性腺苷受体激动剂（N-乙基甲酰氨基）腺苷（NECA）[100 nM（A（1）、A（2A）、A（3））; 10 μ M（A（2B））]和CBF。在存在和不存在腺苷脱氨酶抑制剂，β-氨基-9-（2-羟基-3-壬基）腺嘌呤盐酸盐（10 μ M）的情况下，也用NECA（100 nM或10 μ M）刺激细胞和MTR。ISO和丙卡特罗都以类似于3 Hz的频率刺激野生型和腺苷敲除小鼠的未处理细胞和/或MTR中的CBF。同样，CBF显著增加，与用NECA刺激的BBEC和野生型MTR中的2-3 Hz相似。用NECA刺激的A（1）、A（2A）和A（3）基因敲除小鼠的MTR也显示CBF增加。然而，NECA不能刺激A（2B）基因敲除小鼠MTR中的CBF。为了证实腺苷调节CBF的机制，进行了蛋白激酶活性测定。这些数据表明，NECA刺激的CBF是由cAMP依赖性PKA激活介导的。总的来说，这些数据表明，嘌呤能刺激CBF需要A（2B）腺苷受体激活，可能通过PKA依赖性途径。
Allen-Gipson DS, Blackburn MR, Schneider DJ, Zhang H, Bluitt DL, Jarrell JC, Yanov D, Sisson JH, Wyatt TA. Adenosine activation of A(2B) receptor(s) is essential for stimulated epithelial ciliary motility and clearance. Am J Physiol Lung Cell Mol Physiol 301: L171-L180, 2011. First published May 27, 2011; doi:10.1152/ajplung.00203.2010.-Mucociliary clearance, vital to lung clearance, is dependent on cilia beat frequency (CBF), coordination of cilia, and the maintenance of periciliary fluid. Adenosine, the metabolic breakdown product of ATP, is an important modulator of ciliary motility. However, the contributions of specific adenosine receptors to key airway ciliary motility processes are unclear. We hypothesized that adenosine modulates ciliary motility via activation of its cell surface receptors (A(1), A(2A), A(2B), or A(3)). To test this hypothesis, mouse tracheal rings (MTRs) excised from wild-type and adenosine receptor knockout mice (A(1), A(2A), A(2B), or A(3), respectively), and bovine ciliated bronchial epithelial cells (BBECs) were stimulated with known cilia activators, isoproterenol (ISO; 10 mu M) and/or procaterol (10 mu M), in the presence or absence of 5'-(N-ethylcarboxamido) adenosine (NECA), a nonselective adenosine receptor agonist [100 nM (A(1), A(2A), A(3)); 10 mu M (A(2B))], and CBF was measured. Cells and MTRs were also stimulated with NECA (100 nM or 10 mu M) in the presence and absence of adenosine deaminase inhibitor, erythro-9- (2-hydroxy-3-nonyl) adenine hydrochloride (10 mu M). Both ISO and procaterol stimulated CBF in untreated cells and/or MTRs from both wild-type and adenosine knockout mice by similar to 3 Hz. Likewise, CBF significantly increased similar to 2-3 Hz in BBECs and wild-type MTRs stimulated with NECA. MTRs from A(1), A(2A), and A(3) knockout mice stimulated with NECA also demonstrated an increase in CBF. However, NECA failed to stimulate CBF in MTRs from A(2B) knockout mice. To confirm the mechanism by which adenosine modulates CBF, protein kinase activity assays were conducted. The data revealed that NECA-stimulated CBF is mediated by the activation of cAMP-dependent PKA. Collectively, these data indicate that purinergic stimulation of CBF requires A(2B) adenosine receptor activation, likely via a PKA-dependent pathway.