Antibiotic use is common to treat bacterial infections. It is quite possible that the increased use of antibiotics and the rising incidence of inflammatory bowel diseases (IBD) are linked. This is confirmed by epidemiological studies. However, the mechanism of this link requires a detailed study. Impairment of the intestinal epithelial barrier functioning is an important pathophysiological factor in the IBD development due enhance the immune response to microbial antigens. The aim of the present study was to investigate the effect of antibiotic treatment on the integrity of colonic epithelial barrier. Methods. Male Wistar rats (n = 16, 140–160 g) were treated for 14 days with broadspectrum cephalosporin antibiotic ceftriaxone (300 mg/kg, i.m.) or vehicle. Epithelial permeability was measured on the 15th and 72nd days of the experiment. Evans blue permeation method was used. Rats were anesthetized with urethane (1.1 g/kg, i.p.). Then a laparotomy was performed and the portal vein was catheterized. The rat’s colon was ligated and instilled with 1.5 % Evans blue solution. Blood samples (200 μl) were collected in plastic microtubes containing heparin at 30, 60 and 90 min after intracolonic injection of Evans blue. An equal volume of saline was reinjected after each blood collection.

The blood samples were centrifuged for 5 min (5.000 g, t = 4 °C). The plasmatic absorbance was measured on a spectrophotometer at 620 and 740 nm. Evans blue amount was calculated with following formula: Evans blue content = absorbance at 620 nm – [1.426 × (absorbance at 740 nm) + 0.03]. The colonic epithelial permeability of rats was determined for each rat by the difference between the Evans blue content at time 30, 60 and 90 min after intracolonic injection. Data are presented as M ± SEM. The statistical significance was determined by Student t-test (p < 0.05). Results. A day after ceftriaxone withdrawal (15th day of experiment) amount Evans blue which penetrated to the blood for 30 min (from 30 to 60 min after intracolonic injection) was lower (0.015 ± 0.003 arb. units) compare with the control group of animals (0.028 ± 0.018 arb. units), but these changes were statistically insignificant. For 60 min (from 30 to 90 min after intracolonic injection) the amount of Evans blue which penetrated to the blood was similar for both groups of animals (0,047 ± 0.010 arb. units in ceftriaxone-treated rats vs 0,046 ± 0,012 in control group). Thus ceftriaxone didn't induce significant changes in colonic epithelial permeability day after its withdrawal. At 72nd day of the experiment, average Evans blue amount which penetrated to the blood for 30 min (from 30 to 60 min after intracolonic injection) was increased 2.7-fold (р < 0.05) in ceftriaxone-treated rats in comparison to control group (0.043 ± 0.006 vs 0.016 ± 0.005 arb. units). After 60 min (from 30 to 90 min after intracolonic injection) the amount of Evans blue penetrated to the blood was still increased 2-fold (р < 0.05) in ceftriaxone-treated rats in comparison to control group (0.090 ± 0.004 vs 0.045 ± 0.013 arb. units).

Conclusions. Thus ceftriaxone administration for 14 days increased colon epithelial permeability long-after antibiotic withdrawal. These results indicate that antibiotic therapy might increase susceptibility to inflammatory bowel diseases development by disruption of the intestinal epithelial barrier.

Received 17.11.2016

Keywords: antibiotics, colon, epithelial permeability

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