Secretory effects of kinins on colonic epithelium in relation to prostaglandins released from cells of the lamina propria.

Abstract

1. Sheets of muscle-stripped rat and rabbit colon with epithelium intact or removed were mounted in Ussing-type chambers for recording of transepithelial p.d., resistance and short circuit current (Isc), and measurement by radioimmunoassay (RIA) of the release of prostaglandins into serosal and mucosal bathing solutions. 2. In epithelial-intact preparations prostaglandin E2 (PGE2), PGE1, PGF2 alpha, U46619 and prostacyclin (10(-7)-10(-6) M) caused increases in Isc and transepithelial p.d., in (approximate) descending order of potency. Epithelial-removed preparations did not exhibit any transepithelial p.d. 3. In epithelial-intact preparations, lysyl-bradykinin (LBk) applied serosally but not mucosally caused increased p.d. and release of PGE2 (and to a lesser extent other prostaglandins) into serosal but not mucosal bathing solutions. In epithelial-removed tissues, responsiveness to LBk was maintained, but it did not exhibit 'sidedness', i.e. LBk was effective when applied on either side and PGE2 release occurred into both compartments. 4. Indomethacin and other non steroidal anti-inflammatory drugs (NSAIDs) abolished the LBk-induced p.d. and reduced PGE2 release if applied serosally but not mucosally in epithelial-intact preparations. In epithelial-removed tissues, indomethacin added to either side abolished prostaglandin release into both compartments. 5. Calcium removal from serosal but not mucosal bathing solution (Ca2+-free EGTA Krebs) abolished p.d. generation by LBk in epithelial-intact preparations, and reduced PGE2 release in rabbit but not rat colon. Similarly, in epithelial-removed preparations, calcium removal did not affect kinin-induced PGE2 generation in rat but strongly attenuated it in rabbit colon. 6. We conclude that (i) kinins activate the arachidonate cascade principally by interactions with cells in the subepithelial (lamina propria) layer, rather than with the epithelial cells themselves, (ii) PGE2 contributes substantially to the kinin-induced increase of transepithelial p.d. as a messenger released from kinin-responsive subepithelial cells and acting on the basolateral pole of the epithelial cells, (iii) the apparent sidedness of colonic epithelium in terms of responses to kinins, NSAIDs and calcium removal is due to the barrier properties of the epithelial cell layer, and (iv) there are differences in calcium sequestration and apparent calcium dependence of prostaglandin biosynthesis between rat and rabbit colonic subepithelial cells.