The contributions of subepithelial tissue, mucosa, and mucus gel layer as restraints for oxygen diffusion in rat distal colon in vitro were assessed by comparing oxygen transfer through preparations of isolated submucosa, isolated mucosa with and without the superficial mucus gel layer, and mucosa-submucosa mounted as flat sheets in a diffusion chamber. One side of the chamber was gassed with 95% O2-5% CO2 while the time course of oxygen concentration rise was measured in the continuously stirred opposite side, initially equilibrated with near-zero oxygen solution. The procedure does not affect epithelial viability. Diffusion in isolated mucosa was the same before and after KCN (5 mM) treatment, suggesting that epithelial oxygen consumption does not influence transfer rates. Subepithelial tissue, mucosa, and mucus gel layer are roughly responsible, respectively, for 12%, 56%, and 32% of oxygen diffusive hindrance. Diffusion coefficients range from 13% (mucosa-submucosa) to 54% (isolated submucosa) of that of water. Subepithelial tissue accounts for about 12% of total diffusive restraint.
The aerobic metabolic cost of chloride secretion was studied in rat distal colon isolated mucosa under several conditions by simultaneous measurement of short-circuit current and oxygen consumption under conditions that preserve vectorial ion transport. A low-chloride solution and the presence of bumetanide plus diphenylamine-2-carboxylate reduced short-circuit current by 75% and oxygen consumption by 25%. Ouabain decreased short-circuit current by 93% and oxygen consumption by 32%. Serotonin increased both variables by 59% and 33%, respectively. Bumetanide and diphenylamine-2-carboxylate reduced but did not abolish the effect of serotonin on short-circuit current and oxygen consumption. Changes in short-circuit current and oxygen consumption were linearly correlated under all conditions tested. It is concluded that, in the unstimulated rat distal colon epithelium, chloride secretion accounts for about 75% of ouabain-sensitive short-circuit current and oxygen consumption. Stimulated chloride secretion may demand over 40% of total oxygen consumption.
The distal colon epithelium of rats submitted to chronic hypoxia shows higher short-circuit current (Isc) which, unlike non-hypoxic rat epithelium, has an amiloride-sensitive component despite low serum aldosterone levels. Isc and oxygen consumption (QO2) were simultaneously measured in mucosae from rats submitted to 0.5 atm for 10 days and from control rats in a modified Ussing chamber. Hypoxia increased Isc but not QO2. The slope of the regression line between Isc and QO2 reduction after ouabain addition was decreased in epithelia from hypoxic rats (P = 0.03). Chloride secretion blockade reduced Isc and QO2 in both groups, while sodium channel blockade did so only in the hypoxic group. Dual blockade in hypoxic rat epithelia caused correlated (P = 0.0025) additive decreases in Isc and QO2. Presented results suggest that chronic hypoxia induces an improved coupling between QO2 and electrogenic ion transport, and electrogenic sodium absorption despite low aldosterone levels.
In mucolytic concentration, dithiothreitol modifies ISC and RTE. Oxygen supply from the serosal side is more important to sustain ISC and RTE in biopsy samples. The different effect of hypoxia and Na, K-ATPase blockade on RTE suggests that their depressing effect on ISC involves different mechanisms.
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