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1 Second Division, Department of Internal Medicine, and 2 First Department of Biochemistry, Hamamatsu University School of Medicine, Hamamatsu 431-3192, Japan
ATP induced a biphasic increase in the intracellular Ca2+concentration ([Ca2+]i), an initial spike, and a subsequent plateau in A549 cells. Erythromycin (EM) suppressed the ATP-induced [Ca2+]i spike but only in the presence of extracellular calcium (Ca2+o). It was ineffective against ATP- and UTP-induced inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] formation and UTP-induced [Ca2+]i spike, implying that EM perturbs Ca2+ influx from the extracellular space rather than Ca2+release from intracellular Ca2+ stores via the G protein-phospholipase C-Ins(1,4,5)P3 pathway. A verapamil-sensitive, KCl-induced increase in [Ca2+]i and the Ca2+ influx activated by Ca2+ store depletion were insensitive to EM. 3'-O-(4-benzoylbenzoyl)-ATP evoked an Ca2+o-dependent [Ca2+]i response even in the presence of verapamil or the absence of extracellular Na+, and this response was almost completely abolished by EM pretreatment. RT-PCR analyses revealed that P2X4 as well as P2Y2, P2Y4, and P2Y6 are coexpressed in this cell line. These results suggest that in A549 cells 1) the coexpressed P2X4 and P2Y2/P2Y4 subtypes contribute to the ATP-induced [Ca2+]i spike and 2) EM selectively inhibits Ca2+ influx through the P2X channel. This action of EM may underlie its clinical efficacy in the treatment of airway inflammation.
P2X receptors; P2Y receptors; calcium release; calcium influx; reverse transcriptase-polymerase chain reaction
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