RU2312666C1 - Method for prophylaxis of toxic effect and accumulation of cobalt in experimental animal tissues in modeling chronic poisoning - Google Patents

Abstract

FIELD: experimental biology, toxicology.

SUBSTANCE: method involves every day administration of cobalt chloride into animal stomach by a sound and in a day zeolite as 6% suspension is administrated by the same route. Cobalt chloride and zeolite are administrated simultaneously beginning from the first day of experiment, and irlite-7 is used as zeolite. Method promotes to enhancing elimination of cobalt from body through digestive tract, decreases its content in blood plasma and reduces accumulation of cobalt in liver and kidney tissues.

EFFECT: improved method of prophylaxis.

2 tbl, 1 ex

Description

The invention relates to ecology, medicine, toxicology, experimental biology and can be used to study the mechanisms of excretion of heavy metals, in particular cobalt, from the body.

Increasing volumes of cobalt production, environmental pollution and, as a consequence, the accumulation of cobalt in the tissues of the human body determine the need to create and develop ways to prevent the receipt and enhance the removal of excess metal from the body.

Many detoxicants are known that can remove heavy metals from the body. These include thiol compounds - unitiol, succimer, sodium thiosulfate, various complexones - pentacin, disodium salt of ethylenediaminetetraacetic acid, deferoxamine, Ospenicillamine, etc. (Mashkovsky M.D. Medicines: in 2 volumes. T.2 - Kharkov, "Torsing", 1998. S.212-220). Long-term use of these synthetic drugs can cause various side effects: gastrointestinal tract disorders, allergic reactions, leukopenia, thrombocytopenia, etc. Along with toxins, they can also remove trace elements and vital ions such as iron, calcium, etc., and cause metabolic disorders processes, which complicates their long-term and preventive use.

Currently, the search for substances that contribute to the removal of heavy metals from the body continues. The attention of many researchers is attracted by natural zeolites, unique in their properties, which are aluminosilicates of alkali and alkaline earth metals. Due to their structure and variety of material composition, they possess sorption, ion exchange, and catalytic properties (A.V. Gorbunov et al., 1990; N.I. Petunkin., A.A. Mikhailov et al., 1990; Leach et al., 1990). The sorption properties of zeolites are due to the specific open rigid microporous crystalline lattice of the frame type with a special ability to attach and easily remove water without destroying the structure of the mineral. Zeolites can act as an enterosorbent, promoting the excretion of various compounds from the body, such as urea, creatinine, etc., which leads to a decrease in the proportion of substances excreted by the kidneys (A.D. Gerasev et al., 2001), are capable of adsorbing heavy metal salts have a detoxifying effect on nitrates, nitrites and mycotoxins (B.A. Timofeev et al., 1981; A.F. Kuznetsov et al., 1992). Due to its ion-exchange properties, zeolites can act as sources of trace elements and ions necessary for the body. So, in the works of researchers (V.V. Bayrakov et al., 1985; I.D.Tmenov et al., 2000; I.D.Tmenov, B.A. Dzagurov, 2001) it was shown that due to the presence of zeolites in the diet the coefficient of assimilation of ions of calcium, phosphorus, iron, zinc, etc. increases in animals. Zeolites of different deposits have some differences in physicochemical properties and, consequently, in the degree of positive effect. In 1995, in the mountains of the North Caucasus, on the territory of North Ossetia-Alania, a deposit of natural formations, classified as zeolites, was discovered that surpass in many respects the previously known zeolites. They were called Irlites. Irlit-1 and Irlit-7 are distinguished depending on the chemical composition.

A known method (L.N. Gamko, T.A. Talyzina. Natural zeolite as an absorbent of heavy metals in the body of pigs // Zootechny. - 1997. - No. 2. - P.14-16) removal of lead and cadmium from the body of pigs, using mineral substances - zeolites in the amount of 4-6% of the diet.

The disadvantage of this method is that zeolites are used as feed in the diet, which reduces the accuracy of controlling the dose-dependent effect.

The closest is the method of removing cobalt from the body of experimental animals in case of chronic poisoning, which we took as a prototype (Patent No. 2252765 C1, published May 27, 2005, application No. 2004/01336/14 of January 16, 2004, MKI A61K 33/00, A61P 43 / 00, authors: Albegova NR, Brin VB), which consists in the fact that the animals are first injected with cobalt chloride daily for two months, and then every other day a 6% suspension of zeolite - irlit-1 in the amount of 2.5% of body weight.

The disadvantage of the prototype is that zeolites are used to remove cobalt from the body of experimental animals after creating a model of chronic intoxication, and the prophylactic effect of zeolite has not been investigated.

The invention is aimed at solving the problem of developing a method for preventing the toxic effects of cobalt in chronic poisoning, including the introduction of natural zeolite - irlit-7 simultaneously with intragastric administration of a solution of cobalt chloride.

The solution to this problem provides a significant reduction in the toxic effect of cobalt in chronic poisoning by reducing accumulation and increasing the excretion of heavy metal.

To achieve this technical result, the claimed invention of a method for preventing toxic effects and accumulation of cobalt in tissues in experimental animals when modeling chronic poisoning includes the following essential features: animals are injected with cobalt chloride daily through a tube into the stomach and simultaneously every other day - zeolite - irlite-7 in the form of 6 % Suspension also through a probe into the stomach.

The proposed method differs from the prototype, firstly, the introduction of zeolite Irlit-7, and secondly, the simultaneous introduction (which means the introduction within the same period - 2 months.) Irlit-7 and cobalt chloride reduces the accumulation of cobalt in the body and an increase in its excretion.

The prophylactic administration of natural zeolite - irlit-7 in the form of 6% suspension through a tube into the stomach every other day, simultaneously with the daily administration of a cobalt chloride solution intragastrally, leads to an increase in elimination of cobalt from the body through the gastrointestinal tract, a decrease in its content in blood plasma, and a decrease accumulation in the tissues of the liver and kidneys and a decrease in the pathological effects of heavy metal in experimental animals. The claimed method is effective, cost-effective and easily reproducible.

According to the information available to the authors, the set of essential features characterizing the essence of the claimed invention is not known, which allows us to conclude that the invention meets the criterion of "novelty."

According to the authors, the essence of the claimed invention should not be obvious to specialists from a known level of medicine, since it does not reveal the above possibility of obtaining a method for the prevention of toxic effects and accumulation of cobalt in tissues in experimental animals when modeling chronic poisoning, including daily administration of cobalt chloride through probe into the stomach and in parallel with the first day of the experiment, the introduction of zeolite Irlit-7 in the form of 6% suspension every other day also through the probe into the stomach, which drive ie an increase of cobalt elimination from the body through the gastrointestinal tract, which allows to conclude that the criterion of "inventive step".

The set of essential features characterizing the essence of the invention, in principle, can be repeatedly used in medicine to obtain a result consisting in an effective and easily reproducible method for preventing the toxic effects of cobalt by reducing accumulation and increasing its excretion from the body during chronic poisoning, which allows us to conclude compliance of the invention with the criterion of "industrial applicability".

This method is as follows.

A solution of cobalt chloride is administered daily 1 time per day in an amount of 0.2 mg / kg body weight through a probe into the stomach for two months. At the same time, from the first day of the introduction of cobalt chloride, a suspension of 6% zeolite - irlit-7 in the amount of 2.5% of body weight is injected through the probe into the stomach through a reusable syringe with a specially curved needle on which the probe is put on every other day.

After the experiment (two months), the content of cobalt in the urine and feces was determined in both groups of animals. Rats were killed using thiopental to study the accumulation of cobalt in liver and kidney tissue, to determine its concentration in blood plasma using a plasma (inductively coupled) mass spectrometer ICP MS HP 4500 "HEWLETT - PACKARD".

For histological studies, tissue samples (liver, kidneys) were fixed in 10% neutral formalin, and then subjected to paraffin embedding with the following preparation of sections with a thickness of 7-8 microns. Sections were stained with hematoxylin and eosin. The sections were studied in transmitted light using a Mikmed-1 microscope under a magnification of 80 × 200 × 400. The results were compared with indicators of an intact group of animals and indicators of experimental animals, which were injected only cobalt chloride for two months.

Example 1. Two groups of male rats (No. 1 control and No. 2) of the Wistar line weighing 200-300 g were injected through a tube into the stomach with a solution of cobalt chloride at a dose of 0.2 mg / kg every day for 2 months. Natural zeolite - irlit-7 in the form of 6% suspension in an amount of 2.5% of body weight was injected through a tube into the stomach of rats of group No. 2 every other day. Two months later, a study was conducted of the cobalt content in the tissues of the kidneys and liver, its concentration in blood plasma, in urine and feces, in both groups of animals. Analyzing the data of Tables 1 and 2, it can be noted that with intragastric administration of cobalt chloride, the use of irlite-7 significantly reduces the accumulation of cobalt in the liver tissue (p <0.001) and kidneys (p <0.001), as well as the plasma content (p < 0.02) and urine (p <0.001). An increase in the excretion of cobalt with feces (p <0.001) is noted.

Table 1. The accumulation of cobalt in the tissues of the liver and kidneys with the intragastric administration of cobalt chloride at a dose of 0.2 mg / kg The effect of irlite-7 (M ± m) Groups Kidney, mcg / g Liver, mcg / g background 0.03 ± 0.0002 0.04 ± 0.00003 After 2 months column No. 1 (CoCl ₂ 0.2 mg / kg) 0.2 ± 0.00002 0.16 ± 0.00002 After 2 months column No. 2 (CoCl ₂ 0.2 mg / kg + Irlit7) 0.09 ± 0.00001) * 0.06 ± 0.000005) * ) * - p relative to experience Table 2. The cobalt content in blood plasma, urine and feces with intragastric administration of cobalt chloride at a dose of 0.2 mg / kg The effect of irlite-7 (M ± m) groups Plasma, μg / ml Urine, soda; mcg / ml Cal, mcg / g background 0.001 ± 0.002 0.074 ± 0.010 4.0 ± 0.0031 After 2 months column No. 1 (CoCl ₂ ) 0.012 ± 0.003 0.769 ± 0.146 9.0 ± 0.0003 After 2 months column No. 2 (CoCl ₂ + Irlit-7) 0.004 ± 0.001) * 0.1180.085) * 16.0 ± 0.0018) * ) * - p - relative to experience

Histological examination revealed alterative and necrobiotic processes in all examined organs. In the liver, histostructural changes are presented in the form of focal dystrophic and necrobiotic changes in hepatocytes, hemorrhages in the centers of the liver lobules are noted. In the kidneys, focal dystrophic changes of the glomeruli, edema and small-point hemorrhages, necrobiotic changes of the tubular epithelium are noted. In the group with the prophylactic administration of iritis-7, pathomorphological changes are expressed to a much lesser extent in the form of nephrothelium dystrophy of tubular structures, the appearance of cortical and medullary nephrothelium regeneration sites, signs of active regeneration of endothelium and podocytes. In the liver, moderate fatty degeneration of hepatocytes with signs of their focal regeneration was noted.

It can be seen from the foregoing that the prophylactic administration of zeolite - irlit-7 with the simultaneous administration of cobalt chloride enhances the excretion of cobalt from the body through the gastrointestinal tract, reduces its plasma content, reduces the accumulation of cobalt in the liver and kidney tissues and reduces the pathological effect of heavy metals in experimental animals and, thus, is an effective way to reduce the accumulation and prevent toxic effects of cobalt in chronic poisoning of its compounds eniyami.

Claims (1)

A method for the prevention of toxic effects and accumulation of cobalt in tissues in experimental animals when modeling chronic poisoning, including daily administration of cobalt chloride through a tube into the stomach and every other day a zeolite in the form of 6% suspension also through a tube into the stomach, characterized in that cobalt chloride and zeolite are administered simultaneously from the first day of the experiment, and irlit-7 is used as zeolite.