Intestinal absorption enhanced by unsaturated fatty acids: inhibitory effect of sulfhydryl modifiers.
In vitro absorption of carboxyfluorescein was performed with everted colonic segments of rats. Oleic acid solubilized by a nonionic surfactant HCO-60, but not HCO-60 alone, markedly enhanced the permeation of the otherwise poorly permeant carboxyfluorescein through the colonic mucosa. The effect produced by oleic acid was reduced to different extents by pretreating the mucosa with several SH reagents, with N-ethyl-maleimide being the most effective. The inhibitory effect of N-ethylmaleimide was concentration dependent, with more than 5 mM completely blocking the enhancement of transmucosal permeability by oleic acid. The in vitro effect of N-ethylmaleimide in the absorption experiments was also observed in situ. The inhibitory effects of HgCl2 and iodoacetamide were comparable to that of N-ethylmaleimide, whereas PCMPS, an impermeant SH blocker, had no effect on the enhanced permeation. Various other amino-group modifiers had also no effect on the enhancement. On the other hand, the inhibition was accompanied by a significant reduction in the level of non-protein thiols as well as protein SH groups. Diethyl maleate, which reduced only the non-protein SH level, had no pronounced effect on the oleic acid-induced permeability change. These results suggest that the intact SH group of membrane-associated protein is necessary for the enhanced permeation of carboxyfluorescein elicited by oleic acid.
Murakami
M
M
Department of Biopharmaceutics, Kyoto Pharmaceutical University, Japan.
Takada
K
K
Fujii
T
T
Muranishi
S
S
D000818:Animals; D002368:Castor Oil / Q000031:analogs & derivatives / Q000494:pharmacology; D002463:Cell Membrane Permeability / Q000187:drug effects; D005033:Ethylmaleimide / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D005452:Fluoresceins / Q000378:metabolism; D007408:Intestinal Absorption / Q000187:drug effects*; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000502:physiology*; D019301:Oleic Acid; D009829:Oleic Acids / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D013501:Surface-Active Agents
D005231:Fatty Acids, Unsaturated; D005452:Fluoresceins; D008565:Membrane Proteins; D009829:Oleic Acids; D013439:Sulfhydryl Reagents; D013501:Surface-Active Agents; D019301:Oleic Acid; C024098:6-carboxyfluorescein; C023040:polyethoxylated castor oil; D002368:Castor Oil; D005033:Ethylmaleimide