Genistein stimulates electrogenic Cl−secretion via phosphodiesterase modulation in the mouse jejunum

Abstract

Previously, we demonstrated that genistein stimulated Cl−secretion in the mouse jejunum (Baker MJ and Hamilton KL, Am J Physiol Cell Physiol 287: C1636–C1645, 2004); however, the mode of action of genistein still remains unclear. Here, we examined the activation of Cl−secretion by the modulation of phosphodiesterases (PDEs) by genistein (75 μM) in the mouse jejunum with the Ussing short-circuit current ( Isc) technique. Drugs tested included theophylline (10 mM), a nonspecific PDE inhibitor; 8-methoxymethyl-3-isobutyl-1-methylxanthine (8-MM-IBMX; 100 μM), erythro-9-(2-hydroxyl-3-nonyl)-adenine (EHNA; 40 μM), milrinone (100 μM), and rolipram (40 and 100 μM), which are specific inhibitors of PDE1–PDE4, respectively. Theophylline stimulated a bumetanide-sensitive Isc, indicative of Cl−secretion, and abolished genistein's stimulatory action on Isc. Neither 8-MM-IBMX nor EHNA altered the basal Iscnor did these PDE inhibitors affect the stimulatory action of genistein on the Iscof the mouse jejunum. Rolipram had no effect on basal Isc, but it reduced the genistein-stimulated Isccompared with time-matched control tissues. Milrinone stimulated a concentration-dependent increase in Isc. Bumetanide (10 μM) inhibited 60 ± 4% of milrinone-induced Isc. Pretreating tissues with milrinone prevented genistein from stimulating Isc, and pretreatment with genistein reduced the effect of milrinone on Isc. H89 (50 μM), a PKA inhibitor, reduced the milrinone-stimulated Isc. Likewise, H89 reduced the genistein-stimulated Isc. Here, we demonstrate, for the first time, that genistein activates Cl−secretion of the mouse jejunum via inhibition of a PDE3-dependent pathway.