Enhanced Ca2+ sensitivity in hyperresponsive cultured bronchi is mediated by TNFalpha and NF-kappaB

The mechanical properties and Ca2+ sensitivity of an organ-culture model derived from guinea pig airways have been examined. The cultured explants develop airway hyperresponsiveness to pharmacological agonists after 3-day culture, when compared with fresh and ovalbumin-sensitized tissues. The reactivity of cultured explants is dependent on the presence of the epithelium. They are also sensitive to glucocorticosteroid pretreatments, which neutralize the TNFalpha antibody and NF-kappaB inhibitor. Hence, specific immunostaining of NF-kappaB subunits (p65 and p50) was increased in the nuclear extract of cultured explants. In beta-escin-permeabilized preparations, step-increases in pCa revealed enhanced Ca2+ sensitivity of the contractile apparatus in cultured explants, which was prevented by epithelium removal. Pretreatments of cultured explants with neutralizing TNFalpha antibody and NF-kappaB inhibitor consistently reduced their Ca2+ sensitivity. These findings suggest that AHR developed in this organ culture model may be triggered by an inflammatory process mediated by the TNFalpha and NF-kappaB transcription factor, which results in an increased sensitivity to [Ca2+]i.