Rapid clearance of human insulin-like growth factor binding protein-3 from the rat circulation and cellular localization in liver, kidney and stomach.

Abstract

The gastrointestinal tract represents a major site for the trophic actions of insulin-like growth factors (IGFs), which may be derived in vivo from a large circulating pool. The high capacity binding protein for IGFs in blood is IGF binding protein-3 (IGFBP-3), which is largely complexed with an acid-labile subunit (ALS). However, we and others have shown that IGFBP-3 not complexed with ALS can rapidly leave the circulation, and may carry IGFs to peripheral tissues. In this study we investigated the transfer of recombinant, glycosylated human (h)IGFBP-3 from the rat circulation to the stomach and intestine, compared with liver and kidney. [125I]-labeled IGFBP-3 was administered into the tail vein of conscious male rats, which were killed between 5 min and 2 h later. Blood was taken for the preparation of plasma, and the liver, kidneys, stomach and intestine were removed either for estimation of the associated radioactivity, or fixed for autoradiographic analysis of histological sections. Following injection, [125I]-labeled IGFBP-3 was associated, in part, with a 150 kDa complex in plasma within 10 min when analyzed by gel filtration chromatography. However, 84% of the administered IGFBP-3 had already left the circulation, and 40% of the initial injected dose was accumulated in liver by 5 min, with a further 4% localized in the kidneys. Autoradiographic analysis showed that IGFBP-3 was selectively accumulated within Kupffer cells of the liver, and by the glomeruli and proximal tubules of the kidney. Little radiolabeled IGFBP-3 was recovered from the small intestine, but 14% of the initial injected dose was found within the stomach after 2 h, and a further 12% within the stomach contents. Autoradiographic localization within the stomach showed that the [125I]-labeled IGFBP-3 was primarily associated with the mucosal lining and gastric glands. Separation on sodium dodecyl sulphate polyacrylamide gel electrophoresis showed that the majority of the radioactivity associated with the stomach contents represented small, degraded peptides. These results suggest that while a rapid clearance of IGFBP-3 is achieved by the liver and kidney, a longer term accumulation occurs in the stomach with a luminal secretion. This may represent a delivery system by which circulating IGFs may reach gastric tissue.