BTCI enhances guanylin-induced natriuresis and promotes renal glomerular and tubular effects

Abstract

Guanylin and uroguanylin are small cysteine-rich peptides involved in the regulation of fluid and electrolyte homeostasis through binding and activation of guanylyl cyclases signaling molecules expressed in intestine and kidney. Guanylin is less potent than uroguanylin as a natriuretic agent and is degraded in vitro by chymotrypsin due to unique structural features in the bioactive moiety of the peptide. Thus, the aim of this study was to verify whether or not guanylin is degraded by chymotrypsin-like proteases present in the kidney brush-border membranes. The isolated perfused rat kidney assay was used in this regard. Guanylin (0.2 μM) induced no changes in kidney function. However, when pretreated by the black-eyed pea trypsin and chymotrypsin inhibitor (BTCI - 1.0 μM; guanylin - 0.2 μM) it promoted increases in urine flow (ΔUF of 0.25 ± 0.09 mL.g-1/min, P < 0.05) and Na+ excretion (% Δ ENa+ of 18.20 ± 2.17, P < 0.05). BTCI (1.0 μM) also increased %ENa+ (from 22.8 ± 1.30 to 34.4 ± 3.48, P < 0.05, 90 minutes). Furthermore, BTCI (3.0 μM) induced increases in glomerular filtration rate (GFR; from 0.96 ± 0.02 to 1.28 ± 0.02 mL.g-1/min, P < 0.05, 60 minutes). The present paper strongly suggests that chymotrypsin-like proteases play a role in renal metabolism of guanylin and describes for the first time renal effects induced by a member of the Bowman-Birk family of protease inhibitors.