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Caffeine relaxes smooth muscle through actin depolymerization
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DOI：10.1152/ajplung.00103.2012.png)
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Tazzeo T, Bates G, Roman HN, Lauzon A, Khasnis MD, Eto M, Janssen LJ. Caffeine relaxes smooth muscle through actin depolymerization. Am J Physiol Lung Cell Mol Physiol 303: L334-L342, 2012. First published June 8, 2012; doi:10.1152/ajplung.00103.2012.-Caffeine is sometimes used in cell physiological studies to release internally stored Ca2+. We obtained evidence that caffeine may also act through a different mechanism that has not been previously described and sought to examine this in greater detail. We ruled out a role for phosphodiesterase (PDE) inhibition, since the effect was 1) not reversed by inhibiting PKA or adenylate cyclase; 2) not exacerbated by inhibiting PDE4; and 3) not mimicked by submillimolar caffeine nor theophylline, both of which are sufficient to inhibit PDE. Although caffeine is an agonist of bitter taste receptors, which in turn mediate bronchodilation, its relaxant effect was not mimicked by quinine. After permeabilizing the membrane using beta-escin and depleting the internal Ca2+ store using A23187, we found that 10 mM caffeine reversed tone evoked by direct application of Ca2+, suggesting it functionally antagonizes the contractile apparatus. Using a variety of molecular techniques, we found that caffeine did not affect phosphorylation of myosin light chain (MLC) by MLC kinase, actin-filament motility catalyzed by MLC kinase, phosphorylation of CPI-17 by either protein kinase C or RhoA kinase, nor the activity of MLC-phosphatase. However, we did obtain evidence that caffeine decreased actin filament binding to phosphorylated myosin heads and increased the ratio of globular to filamentous actin in precontracted tissues. We conclude that, in addition to its other non-RyR targets, caffeine also interferes with actin function (decreased binding by myosin, possibly with depolymerization), an effect that should be borne in mind in studies using caffeine to probe excitation-contraction coupling in smooth muscle.
Keyword:
excitation-contraction coupling
Ca2+ handling
airway smooth muscle
ryanodine
asthma
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