19 DIET-INDUCED OBESITY UNIQUELY REGULATES GLUCOSE AND NACL ABSORPTION IN SPRAGUE DAWLEY RAT SMALL INTESTINAL EPITHELIAL CELLS

Abstract

Background Traditional coupled NaCl absorption occurs via the dual operation of Na:H (NHE3) and Cl:HCO3 (DRA/PAT1) exchange in the brush border membrane of villus cells in the mammalian small intestine. In genetic models of obesity (Zucker rat and Tallyho mouse) and humans, coupled NaCl absorption was not affected. However, in these animal models, a novel coupling of Na-glucose co-transport (SGLT1) and Cl:HCO3 exchange was increased, potentially important in the pathogenesis of diabetes and hypertension associated with obesity. Human obesity is both genetic and environmental (e.g. diet mediated). But it is not known whether this novel coupled NaCl absorptive pathway is affected in diet-induced obesity (DIO). Hypothesis Novel coupled NaCl absorption mediated by SGLT1 and DRA/PAT1 is stimulated in DIO. Aim Determine the mechanisms of regulation of SGLT1 and DRA/PAT1 in DIO. Methods Sprague Dawley rats were fed with a high-fat diet (HFD; 60% fat calories) from 12-18 wks of age to induce obesity. Control rats were fed with standard chow. BBM vesicles (BBMV) were prepared from villus cells by Mg precipitation. Phlorizine sensitive, 3H-OMG uptake for SGLT1; amiloride-sensitive, 22Na uptake for NHE3 and DIDS sensitive 36Cl uptake for Cl:HCO3 exchange was performed in villus cell BBMV. Na/KATPase activity was determined as inorganic phosphate release. Western blot studies were performed using rat specific antibodies. Results In intact rat villus cells, SGLT1 uptake was significantly increased in HFD rats (2.4±0.2 nmol/mg protein×2 min in control and 6.4±0.6 in HFD; n=4, p Conclusions In DIO, BBM SGLT1 activity was stimulated in villus cells from HFD rats. This is not due to the altered Na extruding capacity of the cell. Cl:HCO3 exchange was also stimulated in DIO while NHE3 remained unaffected. These data indicate that in DIO, while the traditional coupled NaCl absorption is unaffected, a novel coupled NaCl absorptive pathway, via SGLT1 and DRA/PAT1 is stimulated resulting in enhanced glucose and NaCl absorption which is likely important in the pathophysiology of diabetes and hypertension of obesity.