Actions of hydrogen sulphide on ion transport across rat distal colon

Background and purpose: The aim of this study was to identify the actions of H2S on ion transport across rat distal colon. Experimental approach: Changes in short-circuit current (Isc) induced by the H2S-donor, NaHS, were measured in Ussing chambers. Cytosolic Ca2+ concentration was evaluated using fura-2. Key results: NaHS concentration-dependently induced a change in Isc, that was only partially inhibited by the neurotoxin, tetrodotoxin. Lower concentrations (< 10-3 mol center dot L-1) of NaHS induced a monophasic increase in Isc, whereas higher concentrations induced an additional, secondary fall of Isc, before a third phase when Isc rose again. Blockers of H2S-producing enzymes (expression demonstrated immunohistochemically) decreased basal Isc, suggesting that endogenous production of H2S contributes to spontaneous anion secretion. The positive Isc phases induced by NaHS were due to Cl- secretion as shown by anion substitution and transport inhibitor experiments, whereas the transient negative Isc induced by higher concentrations of the H2S-donor was inhibited by mucosal tetrapentylammonium suggesting a transient K+ secretion. When applied from the serosal side, glibenclamide, an inhibitor of ATP-sensitive K+ channels, and tetrapentylammonium, a blocker of Ca2+-dependent K+ channels, suppressed NaHS-induced Cl- secretion suggesting different types of K+ channels are stimulated by the H2S-donor. NaHS-induced increase in cytosolic Ca2+ concentration was confirmed in isolated, fura-2-loaded colonic crypts. This response was not dependent on extracellular Ca2+, but was inhibited by blockers of intracellular Ca2+ channels present on Ca2+ storage organelles. Conclusions and implications: H2S induces colonic ion secretion by stimulation of apical as well as basolateral epithelial K+ channels.