RU2743803C1 - A method of treating acute experimental pancreatitis by intrarectal administration of a magnesium-containing suppository composition - Google Patents

Abstract

FIELD: medicine, gastroenterology.SUBSTANCE: invention is intended for the treatment of acute pancreatitis as part of the experiment. For this, animals with experimental acute pancreatitis are injected intrarectally with a magnesium-containing composition based on bischofite at the rate of 62.5 mg / kg of magnesium. The injection is carried out once a day for 7 days.EFFECT: method provides decrease in inflammation and fibrosis of the stroma due to magnesium inactivation of the main mechanisms of inflammation development in pancreas.1 cl, 3 dwg

Description

The invention relates to medicine, in particular to experimental medicine, veterinary medicine, and describes a method for treating acute destructive pancreatitis (ADP).

Acute pancreatitis is a poly etiological disease of the pancreas of an inflammatory and destructive nature, the leading links of which are the processes of enzymatic autolysis, necrosis and endogenous infection with involvement of the tissues of the retroperitoneal space, abdominal cavity and extraperitoneal localization systems. [1]

According to the experts of the International Symposium on Acute Pancreatitis (1992) and the IX All-Russian Congress of Surgeons (2020), acute pancreatitis is an acute aseptic inflammation of the pancreas, the basis of which is the process of autoenzymatic necrobiosis, necrosis and endogenous infection with the involvement of the retroperitoneal tissues surrounding it , abdominal cavity and a complex of organ systems of extraperitoneal localization (7).

Despite advances in the diagnosis and treatment of acute destructive pancreatitis (ADP), according to the literature, the incidence is growing from year to year and is still an urgent problem of urgent surgery. The prevalence of acute pancreatitis is up to 389 people per 1 million of the population, every fourth patient develops necrotizing pancreatitis with a total mortality rate of 2.1-15%, and with an infected form of pancreatic necrosis mortality is 30-70% (11) These statistics indicate the need for detailed study of the mechanisms of development of the disease, as well as the search for new methods of pathogenetic treatment.

In order to optimize time and financial costs, a fast, simple, cheap and effective method of treating acute pancreatitis is required.

Due to the fact that in the definition of concepts the primary role is assigned to aseptic inflammation, in which damage to surrounding tissues and distant organs and systems is possible, mechanical trauma of the gland parenchyma was used to model pancreatitis. Trauma always ends with the development of inflammation and brings the model used as close as possible to the clinical conditions of the development of pancreatitis. The leading role in the mechanisms of development of toxemia in acute pancreatitis belongs to pancreatic enzymes: trypsin, lipase, phospholipase A2, lysosomal enzymes that cause oxidative stress, lipid distress, capillary thrombosis, hypoxia, acidosis, endothelial damage.

The primary function of aggression is performed by the pancreatic enzymes trypsin and chymotrypsin, which cause proteolysis of tissue proteins, phospholipase A2 destroys cell membranes, lipase hydrolyzes intracellular triglycerides to fatty acids and combining with calcium leads to lipolytic necrosis in the pancreas and in the retroperitoneal tissue and interstitial connective tissue structures, leading to necrosis.

Pancreatic enzymes activate the components of the kallikrein-kinin system, increase the permeability of the vascular wall, disrupt microcirculation, cause edema, microthrombosis, hypoxia, acidosis.

Tertiary factors of aggression include macrophages, mononuclear cells, neutrophils, against the background of microcirculation disorders, hypoxia and the release of cytokines IL 1, 6.8, TNF, platelet activation factor, nitric oxide. The factors of aggression of the fourth order are cytokines, enzymes, metabolites of various natures, which are formed in the pancreas, intestinal wall, and abdominal cavity. They increase the permeability of the intestinal wall, promote the entry of toxins into the portal veins and systemic circulation, as well as into the lymphatic bed with damage to target organs (8, 9, 10).

Phases of development of acute pancreatitis: 1A - edematous pancreatitis (interstitial). In terms of frequency, it takes 80 - 85% in the structure of the incidence In addition, the development of 1A is possible in the form of necrotizing pancreatitis.

1A - Necrotizing pancreatitis (pancreatic necrosis) is detected in 15-20%) of patients. Both interstitial and necrotic type of acute pancreatitis develops within the first 7 days from the onset of the disease. This period is characterized by the formation of foci of necrosis in the parenchyma and surrounding tissue. Phase 1B is characterized by the body's response to necrosis foci. 11-phase of late sequestration of emerging structures. With rejection, the duct system is depressurized and a pancreatic fistula is formed. To diagnose acute pancreatitis and determine the phase of its development, a number of clinical and laboratory research methods are performed (10). Under experimental conditions, morphological criteria serve as the most convincing evidence, an argument for the development of pancreatitis (4).

The technical task of our invention is the development of a new method of therapy for acute pancreatitis, pathogenetically adequate to the changes developing in the tissues of the gland during its destructive-inflammatory lesions.

The experiments were performed on 15 laboratory rats. Modeling of acute destructive pancreatitis was carried out on sexually mature rats, females of the Wistar line, weighing from 2000 to 400 grams, kept under standard vivarium conditions. When carrying out surgical interventions, the rules of asepsis and antiseptics were observed, as well as the rules provided by the European Commission for the supervision of laboratory and other experiments with the participation of experimental animals of different species. For anesthesia, a solution of rometar and a 2% solution of lidocaine were used (according to the instructions). After treatment of the operating field, an upper midline laparotomy was performed along the white line of the abdomen. The stomach, pancreas and duodenal loop were excreted into the wound. On the surface of the parenchyma of the gland, with a needle from an insulin syringe, to the depth of the needle cut, two parallel incisions 1 cm long were made. After the injury, the organs were immersed in the abdominal cavity, which was tightly sutured with interrupted silk sutures. The wound was treated with 0.02% furacillin solution. The animals were kept in conditions of free access to water and food. After modeling, the animals were divided into two groups: group A and group B. Rats of group B immediately after modeling and for 7 days, once a day, a magnesium-containing suppository drug composition was injected into the rectum at the rate of 62.5 mg / kg / day , in a volume of 0.1-0.18 ml in accordance with the weight of the animal, the volume of the ampoule of its rectum. Suppository magnesium-containing composition based on polymineral bischofite in 0.1 ml of candle mass contains 14 mg of magnesium (5.6). The exposure of the simulation was 7 days. Upon completion of the simulation, blood was obtained for biochemical study of the level of magnesium in blood plasma and erythrocyte mass, and pancreatic tissues were taken for morphological examination.

The positive aspects of the proposed method is the multifaceted role of magnesium in inactivating the mechanisms of inflammation development:

1) A significant role in the pathogenesis of inflammation is assigned to a disorder of peripheral circulation, in particular, ischemia, in the mechanism of formation of which calcium plays, activating angiospasm. Magnesium is a functional calcium antagonist (1).

2) A number of pancreatic enzymes such as phospholipase, protease, lipase are activated by calcium or it refers to a factor that optimizes their activation and is inactivated by magnesium (3). Activated enzymes cause destruction of pancreatic tissue, increasing inflammation.

3) An essential component of inflammation is increased proliferation. Proliferation begins with the formation of TRAMP 5, 6 (3) magnesium channels in the membrane and its entry into the cell. In the cell, it activates more than 300 enzymes of energy production and energy consumption and enhances the formation of ATP.

Thus, magnesium inactivates significant mechanisms of inflammation development.

Examples of specific implementation.

Experience number 1

Rats, Wistar lines of group A and group B, weighing 203 g and 215 g, under anesthesia using rometar and lidocaine (according to the instructions) underwent a midline laparotomy in compliance with the rules of asepsis and antiseptics. A part of the stomach and duodenum with the pancreas were taken out into the operating wound, after which 2 parallel 1 cm incisions were made to the depth of the needle cut from the insulin syringe.After modeling, the organs were immersed back into the abdominal cavity and the anterior abdominal wall was sutured layer by layer with interrupted silk sutures as in group A and group B. A rat of group B, weighing 215 g, immediately after modeling was injected into the rectum 0.1 ml of a magnesium-containing medicinal composition (at the rate of 62.5 mg / kg / day for magnesium). A rat weighing 215 g needs to be injected with 13.43 mg of magnesium, this amount of magnesium is contained in 0.1 ml of a suppository mass. Every day, once a day, a suppository magnesium-containing composition was injected into the rectum for 7 days. At the end of the experiment, after 7 days, the rats were anesthetized again, biological fluids were taken for diagnostic studies (monitoring the level of magnesium in plasma and erythrocyte mass) and tissues were taken for morphological studies.

In the described pathomorphological sample No. 1, group A, after 7 days from the start of modeling, the morphological picture of acute inflammation was determined. In the interlobular stroma, plethora of capillaries and venules, focal edema, focal hemorrhage, inflammatory infiltration by macrophages, eosinophils, neutrophils, and lymphocytes were noted. Focal dystrophic changes in pancreatocytes were revealed: individual cells decreased in size, pronounced basophilia of the cytoplasm was noted, some cells with focal vacuolization of the cytoplasm. In parapancreatic tissue - pronounced plethora of blood vessels, dilated plethora of capillaries and venules, focal edema, in some areas the accumulation of macrophages, lymphocytes.

In pathomorphological sample No. 1, group B, 7 days after the start of modeling against the background of the application of a magnesium-containing composition, the exocrine part of the gland is represented by clearly contoured islets of a rounded shape, formed by endocrinocytes, including full-blooded blood capillaries, focal perivascular weakly expressed infiltration by macrophages, eucatinocytes ... Focal growths of "maturing" granulation tissue in the interlobular stroma.

Comparative analysis of samples from groups A and B shows that the pathomorphosis of acute experimental pancreatitis when using the magnesium composition was characterized by less pronounced vascular and inflammatory reactions, a tendency to the formation of granulation tissue and focal fibrosis of the interlobular stroma.

Experience number 2

Sexually mature rats of the "Wistar" line, weighing 260 g in group A and 410 g in group B under anesthesia underwent upper midline laparotomy, using the rules of asepsis and antiseptics. A part of the stomach and the duodenum with the pancreas were removed into the operating wound, after which 2 parallel 1 cm incisions were made to the depth of the needle cut from the insulin syringe.After modeling, the organs were immersed back into the abdominal cavity and the anterior abdominal wall was sutured layer by layer with interrupted silk sutures. A rat from group B, weighing 410 g, immediately after modeling, was injected intrarectally with a suppository magnesium composition in a volume of 0.18 ml (based on magnesium 25.6 mg per rat weight, this is the content in 0.18 ml of a candle composition), for 7 days one once a day. At the end of the experiment, after 7 days, the rats were anesthetized again, biological fluids were taken for diagnostic studies and tissues were taken for morphological studies.

In the described pathomorphological sample No. 2, group A, 7 days after the start of modeling, the morphological picture of acute inflammation was determined. There was plethora of capillaries and venules, focal edema, significant hemorrhages, focal perivascular inflammatory infiltration by macrophages, eosinophils, neutrophils, and lymphocytes. Focal dystrophic changes in pancreatocytes were revealed: the cells decreased in size, there was an increased basophilia of the cytoplasm, some cells with focal vacuolization of the cytoplasm. In parapancreatic tissue - pronounced vascular reactions: plethora of microvessels - capillaries and venules, edema in places, in some areas accumulation of macrophages, lymphocytes.

In the pathomorphological sample No. 2, group B, 7 days after the start of modeling, against the background of the application of the magnesium-containing composition, the exocrine part of the gland contained exocrinocytes with clear outlines of a rounded shape, in the parapancreatic tissue there was focal perivascular weakly expressed infiltration by macrophages, lymphocytes, single eosinophils. In the interlobular stroma there are focal growths of "maturing" granulation tissue.

Comparative analysis of samples from groups A and B shows that the pathomorphosis of acute experimental pancreatitis when using the magnesium composition was characterized by less pronounced vascular and inflammatory reactions, a tendency to the formation of granulation tissue and focal fibrosis of the stroma.

Example 3.

Rats, Wistar lines of group A and group B, weighing 207 g and 305 g, respectively, under anesthesia using rometar and lidocaine (according to the instructions) underwent median laparotomy in compliance with the rules of asepsis and antiseptics. A part of the stomach and duodenum with the pancreas were taken out into the operating wound, after which 2 parallel 1 cm incisions were made to the depth of the needle cut from the insulin syringe.After modeling, the organs were immersed back into the abdominal cavity and the anterior abdominal wall was sutured layer by layer with interrupted silk sutures as in group A and group B. A rat of group B, immediately after modeling, 0.14 ml of a magnesium-containing medicinal composition was injected into the rectum (the magnesium content based on the weight of the rat is 19.06 mg, it is contained in 0.14 ml of the candle mass) ... Every day, once a day, a suppository magnesium-containing composition was injected into the rectum for 7 days. At the end of the experiment, after 7 days, the rats were anesthetized again, biological fluids were taken for diagnostic studies (monitoring the level of magnesium in plasma and erythrocyte mass) and tissues were taken for morphological studies.

In the described pathomorphological sample No. 1, group A, after 7 days from the start of modeling, the morphological picture of acute inflammation was determined. When the tissues were stained with hematoxylin-eosin, moderately pronounced plethora of interstitial vessels was noted, with focal dislocation of the vessel walls, moderately pronounced edema and inflammatory infiltration by eosinophils, macrophages, and lymphocytes. There are focal dystrophic changes in pancreatic cells, in a number of areas lymphocytes and macrophages of the inflammatory infiltrate are closely located to the basement membrane of acinar cells. In some areas - focal leukopedesis. In the parapancreatic tissue, there is a pronounced plethora of blood vessels, focal diffuse hemorrhages, dissociation of the walls of blood vessels with moderate edema, moderate infiltration by eosinophils, macrophages, and lymphocytes.

In the pathomorphological sample No. 3, group B, 7 days after the start of modeling, against the background of the application of a magnesium-containing composition, the exocrine part of the gland contained exocrinocytes with clear rounded outlines. A weakly expressed inflammatory infiltration of the interlobular stroma by macrophages, lymphocytes, eosinophils was revealed; in the parapancreatic tissue, focal perivascular weakly expressed infiltration by macrophages, lymphocytes with single eosinophils was detected. In the interlobular stroma there are focal growths of "maturing" granulation tissue.

Comparative analysis of samples from groups A and B shows that the pathomorphosis of acute experimental pancreatitis when using the magnesium composition was characterized by less pronounced vascular and inflammatory reactions, a tendency to the formation of granulation tissue and focal fibrosis of the stroma.

Thus, this study allows us to conclude that the use of a suppository magnesium-containing composition leads to a decrease in the intensity of inflammation in the tissues of the pancreas and fibrosis of the stroma. The results obtained make it possible to recommend the use of a magnesium-containing composition for the treatment of acute pancreatitis in veterinary medicine and medicine.

Claims (1)

A method of treating acute experimental pancreatitis by intrarectal administration of a magnesium-containing suppository composition based on polymineral bischofite in a volume of 0.1-0.18 ml, containing an estimated magnesium content: 62.5 mg / kg / day, frequency of application once a day, duration of treatment 7 days.