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ECPB 2014, 68(4): 25–29
Experimental physiology and biochemistry

Determination of oxyproline areas intestinal anastomosis after resection of the small intestine in terms of peritonitis

BLIKHARSKYI YU., MAZUR YU., FEDUSHCHAK N, KRAMARENKO S.
Abstract

As a result of the studies risk factors for failure of anastomosis were formulated. It is impossible to differentiate the impact of some factors on the actions of others. That is why, to understand the process of healing intestinal wounds, the greatest weight are experimental works. The aim of our study was to determine the concentration of hydroxyproline in the area of the anastomosis in a case of peritonitis, in experiments on rabbits.

The experiment was performed on sexually mature males rabbits in the operating conditions of the vivarium and laboratory of Toxicology and Analytical Chemistry. Surgical intervention was carried out under the European Convention for the Protection of Vertebrate Animals used for experimental and other scientific purposes (Strasbourg, 1986), the Law of Ukraine "On protection of animals from cruelty" and were approved due to the local Ethics Committee.

In the operating room of vivarium, subject to the rules of aseptic and antiseptic, medium median laparotomnoy access was used. Intestinal sewed end-to-end single node sero-muscular sutures using microsurgical instruments and atraumatic suture material prolen 6,0. Peritonitis modeled by enterotomy of devascularization section of the small intestine at a distance of 15 cm before anastomosis. Laparotomy wound sewn seams single node through the all layers. Definition of the oxyproline in segment of the small intestine with anastomosis was done on second day after operation. A part of intestine crushed in a mortar. 0.1–0.4 g of tissue were taken and homogenized with 3 ml of distilled water in homogenizer HG-15A-Set. To blenders added 5 ml of concentrated hydrochloric acid, closed tightly in a bottle and heated to 120°C for 16 h. After 16 hours the contents of the vial were transferred to a porcelain cup and evaporated liquid from the test material in a water bath. The dry residue was dissolved in distilled water and the total volume of the solution was adjusted to 10 ml and centrifuged in a centrifuge Engelsdorf-Leipzig Janetzki for 10 min. at a speed of 5000 rev/ min. 4 ml of the supernatant brought into the test tube, added 2 ml of 1% solution of chloramine B, stirred and left for 20 minutes. After that 3.15 M solution of perchloric acid was added to 2 mL and left for 5 minutes, adding 2 ml Ehrlich reagent and heated keeping the temperature of 60°C for 20 minutes. We used spectrophotometry for determination of hydroxyproline on СФ-46 at a wavelength of 480 nm.

Based on previous studies, the concentration of hydroxyproline intact wall of the small intestine ranged from 33.02 to 38.24 mg/100 g. On the third day after the operetion hydroxyproline compounds decreased slightly from 29.02 to 36.23 mg/100 g. Hydroxyproline concentration of area of intestinal anastomosis in peritonitis is from 22.03 to 26.85 mg / 100 g. In the case of peritonitis observed decrease oxyproline concentration compared to intact intestine and anastomosis on the third day after surgery. Peritonitis increases collagen destruction of area of intestinal wound. It makes us look for ways to protect the anastomosis in terms of peritonitis.

Keywords: hydroxyproline, intestinal wound, peritonitis

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