RU2286608C1 - Method for modeling acute necrotic pancreatitis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves subjecting adult rat to midline laparotomy under ether anesthesia, exposing the pancreas and introducing 0.2 ml of complex preparation having 10% aqueous lysine solution and 10% aqueous Triton X-100 solution taken in equal volumes.

EFFECT: irreversible cytoplasmic pancreas cell membranes destruction.

4 dwg

Description

The invention relates to medicine, namely to the field of reproduction of human diseases in an experiment, and can be used to model pancreatic necrosis with the subsequent study of the pathogenesis of this disease and its treatment. The inventive method relates to traumatic models of acute pancreatitis, reproducible using chemical agents.

V.O.Popov, Yu.S. Vinnik and M.I. Gulman in his monograph on modeling acute pancreatitis (Popov V.O., Vinnik Yu.S., Gulman M.I. Acute pancreatitis: pathogenetic correction in experimental conditions . - Krasnoyarsk: GUPP Sibir, 2000, pp. 10-15) provide data on the creation of experimental acute pancreatitis using various chemical agents: physiological saline (Berdavgetov B.A., 1972; Zhandarov N.I., 1983), potassium permanganate (A. Shalimov et al., 1983), sodium taurocholate (Gankish PG, 1974; Rosato EF et al., 1972), pilocarpine (Perelman A.D., 1979), ski Idar (Pigarevsky JO, 1978).

To date, there are many ways to simulate acute inflammation of the pancreas, including using chemical agents, but only a small part of them are simple and convenient reproducibility, as well as high efficiency.

To reproduce pancreatic necrosis in rats, a method of exposure to rat pancreatic tissue by low-frequency ultrasound is known (RF patent No. 2195710, G 09 B 23/28, publication December 27, 2002). Known surgical methods for modeling acute pancreatitis in rats (RF patent No. 2236709, G 09 B 23/28, published 09.20.2004; RF patent No. 2230373, G 09 B 23/28, published 06.10.2004). The disadvantages of the above experimental models is the high complexity and fairly low reproducibility.

The prototype of the proposed method for modeling pancreatic necrosis, the method of modeling pancreatitis using triton X-100, proposed by E.S. Guliants and co-authors, USSR copyright certificate No. 1337152, published July 30, 1987.

During the development of the claimed experimental model, two important tasks were posed: the model should be distinguished by its simplicity and high reproducibility of acute necrotic pancreatitis, which is as close as possible to that of a person.

The problem is achieved in that a dose of 0.2 ml of a complex consisting of a 1% aqueous solution of X-100 triton and a 10% aqueous solution of the basic amino acid lysine taken in equal volumes is injected into the pancreas of an adult rat.

The method is illustrated in figure 1, which shows acute interstitial inflammation of the pancreas with foci of pancreatic necrosis; figure 2 - formed pancreatic necrosis; figure 3 - edema and neutrophilic infiltration of interlobular connective tissue (fragment "a" of figure 2, magnification × 400); figure 4 - whole and destroyed neutrophils in the area of the leukocyte shaft (fragment "b" of figure 2, magnification × 400).

The simulation method is as follows.

A rat weighing 150-250 g under ether anesthesia undergoes median laparotomy, the pancreas is secreted, and a dose of 0.2 ml of a 10% aqueous solution of lysine and 1% aqueous solution of X-100 newt taken in equal volumes is administered. The above dose is not administered simultaneously, but gradually, chopping the pancreas along its entire length, which also contributes to a higher reproducibility of the model. Then the surgical wound is sutured in layers tightly.

The reproducibility of the model increases significantly due to the fact that the main amino acid lysine is added to the X-100 triton, which contributes to the formation of perforations in the biological membranes of pancreatic cells, which is confirmed experimentally. Thus, the combination of Triton X-100 with lysine promotes loosening of the cytoplasmic membranes of pancreatocytes and ultimately leads to an irreversible destructive process in the cell membranes. The reproducibility of the simulated disease has become almost 100% and as close as possible to the pathogenesis of acute necrotic pancreatitis in humans, which allows us to adequately study the pathogenesis of this disease, as well as to study new methods for its treatment. The advantage of this method is its simplicity and versatility, since using the above combination of chemicals it is possible to reproduce necrotic and cytolytic processes in various biological systems.

Changes in pancreatic tissue, interpreted as acute pancreatitis, are confirmed by histological examination. For histological examination, pieces of the pancreas of experimental rats were fixed in a 10% solution of neutral formalin. Paraffin sections were stained with hematoxylin-eosin. Material sampling was carried out 24 and 72 hours after the introduction of 10% lysine solution and 1% aqueous solution of X-100 newt into the pancreatic tissue.

Histological studies showed that after a day in the pancreas dystrophic changes in certain groups of acini are noted (Fig. 1). Three days later, an increase in the severity of dystrophic changes in acinus cells, polymorphonuclear infiltration and interstitial edema is observed in the pancreas of experimental rats, extensive zones of necrosis appear, a leukocyte shaft forms, which is typical for severe acute necrotizing pancreatitis - pancreatic necrosis, which is illustrated in the picture Fig.2-4).

Claims (1)

A method for simulating acute necrotic pancreatitis, including destruction of rat pancreas tissue by Triton X-100, characterized in that a dose of 0.2 ml of a complex consisting of a 1% aqueous solution of X-100 Triton and 10% is injected into the pancreas of an adult rat aqueous solution of the amino acid lysine, taken in equal volumes, receiving formed pancreatic necrosis within 24-72 hours.

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