LTD4 induces hyperresponsiveness to histamine in bovine airway smooth muscle: role of SR-ATPase Ca2+ pump and tyrosine kinase

Airway hyperresponsiveness is a key feature of asthma, but its mechanisms remain poorly understood. Leukotriene D-4 (LTD4) is one of the few molecules capable of producing airway hyperresponsiveness. In this study, LTD4, but not leukotriene C-4 (LTC4), produced a leftward displacement of the concentration-response curve to histamine in bovine airway smooth muscle strips. Neither LTC4 nor LTD4 modified the concentration-response curve to carbachol. In simultaneous measurements of intracellular Ca2+ ([Ca2+](i)) and contraction, histamine or carbachol produced a transient Ca2+ peak followed by a plateau, along with a contraction. LTD4 increased the histamine-induced transient Ca2+ peak and contraction but did not modify responses to carbachol. Enhanced responses to histamine induced by LTD4 were not modified by staurosporine or chelerythrine but were abolished by genistein. Western blot showed that carbachol, but not histamine, caused intense phosphorylation of extracellular signal-regulated kinase 1/2 and that LTD4 significantly enhanced the phosphorylation induced by histamine, but not by carbachol. L-type Ca2+ channel participation in the hyperresponsiveness to histamine was discarded because LTD4 did not modify the [Ca2+](i) changes induced by KCl. In tracheal myocytes, LTD4 enhanced the transient Ca2+ peak induced by histamine ( but not by carbachol) and the sarcoplasmic reticulum (SR) Ca2+ refilling. Genistein abolished this last LTD4 effect. Partial blockade of the SR-ATPase Ca2+ pump with cyclopiazonic acid reduced the Ca2+ transient peak induced by histamine but not by carbachol. These results suggested that LTD4 induces hyperresponsiveness to histamine through activation of the tyrosine kinase pathway and an increasing SR-ATPase Ca2+ pump activity. L-type Ca2+ channels seemed not to be involved in this phenomenon.