Thiamine Transport Across the Rat Intestine: I. Normal characteristics

The characteristics of normal thiamine transport across the intestine were studied in rats using intact intestinal loops and everted jejunal segments. In vivo studies with [³⁵S]-thiamine hydrochloride revealed, in all segments of small intestine, saturation kinetics for low thiamine concentrations (0.06 to 1.5 μM), but a linear relationship between high concentrations (2 to 560 μM) and absorption. Moreover, in vitro studies of net transmural flux using everted jejunal sacs demonstrated movement of [¹⁴C]-thiamine hydrochloride against a concentration gradient only when low, but not when high, thiamine concentration was used, so that the serosal to mucosal ratio became significantly greater than the initial value of one. Pyrithiamine, 2 μM, dinitrophenol, 200 μM, norethylmaleimide, 100 μM, and ouabain, 100 μM, reduced the net transmural flux of 0.2 μM thiamine. In contrast, these inhibitors had no effect on 20 μM thiamine. When unidirectional flux across the jejunum was measured, saturation kinetics was again demonstrated for low thiamine concentrations. This phenomenon, however, was abolished by the addition of pyrithiamine, which exerted competitive inhibition on thiamine absorption. Anoxia and sodium lack reduced intestinal uptake of 0.5 μM thiamine to 58% and 74% of normal, respectively, but did not affect uptake of 50 μM thiamine. Lowering the temperature led to a fall in thiamine uptake, the reduction being more marked with low thiamine concentrations (Q₁₀, 1.648) than with high concentration (Q₁₀, 1.127). Stirring of the water layer reduced K_m to 59% of unstirred value, while the V_max and permeability coefficient remained unchanged. Finally, movement of low concentration thiamine against an electrical gradient was observed under conditions of electrical short circuiting and zero potential difference. In contrast, no such effect was seen with high concentrations. These studies suggest that there exists in the rat a dual system of intestinal thiamine transport. At low concentrations, thiamine is absorbed by an active process; at high concentrations, transport across the intestine is largely a passive movement.