Method of simulation of chemical gastritis in experiment

FIELD: medicine.

SUBSTANCE: for simulation of chemical gastritis in nonlinear white rats a mixture of 200 mcl of 5% solution of ascorbic acid and nonsteroidal antiinflammatory preparation Ketorol are administered orally during 21 days at the rate of 5 mg/kg 1 hour prior to feeding once a day.

EFFECT: adequate reproduction of the model.

2 ex

 

The invention relates to medicine, namely to gastroenterology, pathological physiology and experimental medicine and can be used to study the pathogenesis of chemical gastritis and preclinical efficacy studies of new methods of treatment of this pathology.

Today there are several options for modeling chemical gastritis in the experiment. The closest analogue is the way of modeling the chemical gastritis in rats male Wistar by simultaneous introduction of animals nonsteroidal anti-inflammatory drugs and medical bile (Knyshov CENTURIES the Influence of boron mineral water on the status of processes of lipid peroxidation and antioxidant factors of protection in experimental gastroduodenitis // Questions of balneology, physiotherapy and exercise therapy. - 2002. No. 2. - P.34-36).

The disadvantage of this method is the absence of such pathognomonic for chemical gastritis and histological signs as hyperplasia pit epithelium with local depletion of mucin, hypertrophy of the smooth muscle with the least pronounced swelling and inflammation in its plate mucosa, increase in its plate gastric mucosa in the number of smooth muscle cells (Sobala, G.M. Reflux gastritis in the intact stomach / Sobala G.M., King, R.F., Axon, A.T., Dixon M.F. // J. Clin. Pahol. - 1990, Apr. - Vol.43. - 34. - P.303-306 Compton K.K. Gastritis: new pathological classification and diagnosis / K. Compton // ROS. Journe. Gastroenterol., gepatol., coloproctol. - 1998. No. 3. - P.84-80). On the contrary, the author notes the emergence of a diffuse lymphocytic-histiocytomas and plasmocytomas inflammatory infiltration own plates mucous stasis and small hemorrhages, cubic transformation of epithelial and glandular epithelium, which is not typical for chemical gastritis. Therefore, in modern conditions of development of optimal, approximate to the real conditions of option modeling of chemical gastritis is an important task.

Effect: create a model, which reflects the morphological picture of the destruction of the gastric mucosa in chemical gastritis in humans.

This task is achieved by oral administration of nonlinear white rats for 21 days of a mixture of 200 μl of 5% solution of ascorbic acid and non-steroidal anti-inflammatory drug Ketorol®" ("Dr. Reddy with laboratories Ltd., India, haiderabad, active substance - Ketorolac tromethamine) at a rate of 5 mg/kg for 1 h before feeding 1 time per day.

The method is as follows. Nonlinear white rats (males and females) within 21 days orally administered mixture of 200 μl of 5% of the races is the thief of ascorbic acid and non-steroidal anti-inflammatory drug Ketorol rate of 5 mg/kg for 1 h before feeding 1 time per day. At the end of the experiment animals slaughtered under ether anesthesia with observance of the rules of euthanasia, the material (stomach wall) fixed in 10% buffered by Lilly (pH 7,2) formalin. Using conventional techniques, prepare histological specimens, which are revealed typical histological signs of chemical gastritis: the pit epithelium hyperplasia with local depletion of mucin, the increase in the number of tuck-in (parietal) cells in the bottom of the glands of the mucous membrane of the stomach pyloric hypertrophy of muscle plate of the gastric mucosa, increase in its plate gastric mucosa in the number of smooth muscle cells, edema own plates mucous membrane.

The experiment was performed on 30 animals - male and female outbred white rats four months of age contained in standard vivarium conditions. All experiments were conducted in accordance with "the Rules of work with the use of experimental animals" (Annex to order of the Ministry of health of the USSR from 12.08.1977, N 755) and the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes" dated March 18, 1986

Animals were divided into 2 experimental groups, the composition of the groups was as follows:

Group I - control (INTA the local animals), n=15;

Group II - animals treated for 21 days orally 1 time a day for 1 hour before feeding a mixture of 200 μl of 5% solution of ascorbic acid and non-steroidal anti-inflammatory drug Ketorol®" ("Dr. Reddy with laboratories Ltd., India) at a rate of 5 mg/kg, n=15.

On the 22nd day from the beginning of the experiment, animals were taken from experiment by transection of the spinal cord under ether anesthesia with observance of the rules of euthanasia. When conducting autopsy histological samples obtained from different parts of the rat stomach (esophageal, cardiac, pyloric departments)were fixed in 10% buffered by Lilly formalin (pH of 7.2), obezvozhivani in ascending alcohols fortress, were placed in paraffin, and sections (3-4 μm) were obtained on a rotary microtome, were applied to the glass with adhesive coating (egg albumin), were stained with hematoxylin and eosin and pikrofusin by van gieson.

When conducting a histological study in all animals treated within 21 days of the mixture of 5% ascorbic acid and non-steroidal anti-inflammatory drug Ketorol, noted the appearance of histological signs pathognomonic for chemical gastritis: the pit epithelium hyperplasia with local depletion of mucin, reducing the number of main cells in the bottom of the glands of the mucosa of the pyloric stomach, hypertro the Oia muscle plate of the gastric mucosa, the increase in own plate of gastric mucosa in the number of smooth muscle cells and edema.

Examples of specific performance

Example 1. Male white rats weighing 210 g for 21 days orally 1 hour before feeding 1 time per day was administered a mixture of 5% solution of ascorbic acid and non-steroidal anti-inflammatory drug Ketorol rate of 5 mg/kg On the 22nd day the animal was killed under ether anesthesia by transection of the spinal cord, performed histological examination of the stomach wall.

In the study of histological specimens in transmitted light of the light microscope revealed visible plethora of vessels and edema own plates mucosal and submucosal basis gastric esophageal, cardiac, fundic and pyloric regions. In the glandular mucosa of the stomach was observed pit epithelium hyperplasia, a local decrease in the number of mucous cells, the increase in the number of parietal cells in the bottom of the glands of the gastric mucosa, muscle hypertrophy plate mucosa, the emergence of the private record of the mucous membrane of the stomach of a large number of smooth myocytes. Identified changes are consistent with the histological picture of NSAID gastropathy.

Example 2. The female white rats weighing 178 g for 21 days PE is given orally 1 hour before feeding 1 time per day was administered a mixture of 5% solution of ascorbic acid and non-steroidal anti-inflammatory drug Ketorol®at the rate of 5 mg/kg On the 22nd day the animal was killed under ether anesthesia by transection of the spinal cord in compliance with the rules of euthanasia performed histological examination of the stomach wall.

Microscopic examination of histological samples noted marked plethora of vessels and edema own plates mucosal and submucosal basis gastric esophageal, cardiac, fundic and pyloric regions. In the glandular portion of the gastric mucosa revealed hyperplasia pit epithelium, a local decrease in the number of mucous cells, reducing the number of main cells due to the predominance of tuck-cells in the bottom of the glands of the mucous membrane of the stomach pyloric hypertrophy in the muscle plate of the mucous membrane, the emergence of the private record of the mucous membrane of the stomach of a large number of smooth muscle cells.

Thus, this animal daily oral administration of 5% solution of ascorbic acid in combination with nonsteroidal anti-inflammatory drug ("Ketorol®") has led to the development of chemical gastritis, which is confirmed by histological examination.

The positive effect from the use of the proposed method is to create experimental models of chemical gastritis, with 100%reproducibility and corresponding histological picture of the chemical gastritis, observed in humans.

A method for simulating chemical gastritis in the experiment by oral administration to rats nonsteroidal anti-inflammatory drug, wherein the nonlinear white rats for 21 days to enter a mixture of 200 μl of a 5%solution of ascorbic acid and non-steroidal anti-inflammatory drug Ketorol rate of 5 mg/kg for 1 h before feeding, 1 time per day.



 

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