RU2329793C1 - Method of animal oxidative stress treatment and prevention - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science. Method includes permanent oral introduction of microelement preparation "Helavite" in preventive daily dosage 0.01-0.02 ml per 1 kg of animal live weight mixed with feed, treatment daily dosage is 0.05-0.07 ml per 1 kg of live weight within 4 weeks followed with change to preventive dosage. Method enables to normalise erythropoiesis, to increase activity of catalase and superoxide dismutase that leads to decreased lipid peroxidation content in animal blood serum.

EFFECT: development of effective method of oxidative stress treatment and prevention in veterinary science.

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Description

The invention relates to animal husbandry and veterinary medicine and can be used for the prevention and treatment of oxidative stress in animals.

The initial stage of the development of oxidative stress is the excessive formation of highly active free radical forms of oxygen. The resulting free radical forms of oxygen act on phospholipids, more precisely on unsaturated fatty acids, and they are subjected to peroxidation. During this oxidation, free radical forms of these acids are formed with damaging properties and toxic oxidation products. As a result, cell structures are destroyed up to cell death.

The participation of free radicals in the pathogenesis of many diseases has been proved - shock of various origins, impaired cerebral, coronary and peripheral circulation, diabetes mellitus, inflammatory and degenerative diseases of the musculoskeletal system, respiratory tract diseases, oncological pathology, premature aging, etc.

The process of free radical oxidation would develop uncontrollably if there were no antioxidants in the cell elements that counteract its course. This group of substances includes both antioxidant enzymes (superoxide dismutase, glutathione peroxidase, catalase) and non-enzymatic substances (vitamins A, D, E), which belong to the class of quenching (chain-breaking) antioxidants.

Superoxide dismutase (SOD) is a metal-containing protein that is present in the cell in two forms: a cytosolic one, consisting of two similar subunits containing one copper and zinc ion and a mitochondrial one containing manganese ions. SOD has a wide range of activity (pH from 4.8-10.2) and the main biological function of this enzyme is to catalyze the reaction of the formation of hydrogen peroxide from two superoxide anion radicals.

Catalase is a hemoprotein containing four heme groups. This enzyme catalyzes the decomposition of hydrogen peroxide into water and molecular oxygen.

The composition of this enzyme includes such a trace element as iron.

Glutathione peroxidase is a selenium-containing enzyme that catalyzes the conversion of hydrogen peroxide and organic peroxides to hydro compounds, which can be further metabolized by cellular systems.

Vitamin E regulates the intensity of free radical reactions in cells, interrupts the oxidation chain. Vitamin C is an “interceptor" of free radicals, prevents the accumulation of peroxides. Vitamin A can be a “trap” of free radicals, singlet oxygen, as well as a sulfhydryl reducing agent.

Strengthening the processes of free radical oxidation accompanies diseases with severe stress, inflammatory reactions, hypoxic and ischemic conditions, an increased content of peroxides creates a negative contribution to the progression of many pathological conditions.

In veterinary practice, various antioxidants, mainly containing selenium and vitamin preparations, are used to prevent oxidative stress.

A known method for the prevention of poisoning of the body of dry cows with nitrates, including the introduction of an intramuscular injection of 1% sodium selenite solution into the animals 60-20 days before calving. At the same time, cases of animal disease with mastitis and endometritis are reduced (RU 2084227, class A61K 33/04, publ. 07.20.97, Bull. No. 20). The method includes the introduction of only selenium, which reduces the effectiveness of the method.

Also known is a medicine for the treatment and prevention of diseases caused by selenium deficiency in the body of farm animals, containing sodium selenite and barium selenite, aminoacetic acid and liquid paraffin (RU 2200013 C2, class A61K 33/04, published March 10, 2003. Bull. No. 7). The drug has low toxicity and a long therapeutic effect. However, the method requires preheating the suspension to 38-40 ° C and intramuscular injection. The drug contains only one selenium, which reduces its effectiveness.

A known method of increasing the antioxidant potential of selenium-containing aqueous solutions (RU 2277915 C2, class A61K 33/04, publ. 06/20/2006, Bull. No. 17). The method relates to the use of a selenite-containing solution stabilized with pharmaceutically acceptable acids selected from citric, acetic, malic and carbonic acids, fruit acids and mixtures thereof for the manufacture of an external medicine for the prevention or treatment of infections caused by the herpes virus, in particular infections of vesicular lichen or age spots. The medicine is a gel or emulsion with a pH of less than 5.0. The method has limited use only for external use and its use in animals is unknown.

Known trace element preparation for animals - chelavit (RU 2227800 C1, publ. 06/20/2006. Bull. No. 17, A23K 1/175, A61K 31/295), consisting of the following components, wt.%: 2Na- or 2K-salt ethylenediamine -N, N ¹ -diuccinic acid 15.0-35.0; Na- or K-salt of amino acids 2.0-10.0; iron (III) 0.6-3.0; manganese (II) 0.5-2.5; copper (II) 0.05-0.25: zinc (II) 0.3-2.5; cobalt (II) 0.005-0.05; selenium (IV) 0.01-0.03; iodine (I) 0.03-0.08; water the rest. In this preparation, trace elements are in a balanced state in the form of chelates. The use of the drug enhances the reproductive ability of animals, stimulates the growth, development and productivity of agricultural and domestic animals. However, the use of this composition to prevent oxidative stress in animals is not known.

There is also known a method of normalizing immunobiochemical homeostasis of cows in the pre- and postpartum periods, including the combined use of a 0.5% solution of sodium selenite and active substances such as tetravite and succinic acid (RU 2252767 C1, class A61K 33/04, publ. 27.05. 2005. Bull. No. 15). The method includes intramuscular administration to cows at certain time periods of 0.5% sodium selenite, tetravit solution (vitamins A, D, E, F) and oral administration of succinic acid according to a specific scheme. The method provides an effective correction of the main indicators of homeostasis in cows after calving. However, the application of the method for pets is very difficult, because the method requires repeated intramuscular injections and complex therapy under the supervision of a doctor.

The objective of the invention is to increase the effectiveness of the method of treatment and prevention of oxidative stress in animals, as well as its simplification.

To solve the problem in a method of treating and preventing oxidative stress in animals using a drug containing selenium and active substances, the microelement preparation Helavit is used as a medicine according to the invention for prophylaxis in a daily dose of 0.01-0.02 ml per 1 kg of live weight of the animal orally with food with constant use, for treatment - in a daily dose of 0.05-0.07 ml per 1 kg of live weight for 4 weeks, followed by a transition to a prophylactic dose. The difference of this microelement preparation from others is the chelated form of the complex of Fe, Cu, Mn, Zn, Co, Se and J with succinic acid derivatives and a number of essential amino acids. Due to the coordination of metals with amino acids, the antagonism between various trace elements is excluded, the pharmacological effect of biochemically active trace elements involved in hematopoiesis, immunobiochemical reactions is enhanced. For the prevention of oxidative stress, a prophylactic dosage of 0.01-0.02 ml of the drug per 1 kg of animal body weight per day is necessary. In conditions accompanied by active oxidative stress (pregnancy, aging, intensive growth, etc.), as well as in pathological conditions, additional antioxidant therapy is required in a daily dosage of 0.05-0.07 ml of the Helavit preparation per 1 kg of live weight of the animal in for 4 weeks, followed by a transition to a prophylactic dosage.

With a preventive dosage of less than 0.01 ml per 1 kg of live weight, the effectiveness of the method decreases, and with therapeutic dosages of more than 0.07 ml per 1 kg of live weight, an excess of biologically active selenium can enter the body of the animal.

The effectiveness of the method of treatment and prevention of oxidative stress (free radical pathology) in animals can be illustrated by the following examples.

Example 1

The study of the effectiveness of the proposed method was carried out on a group of clinically healthy dogs (n = 15) large breeds (German and Central Asian shepherd dogs) aged 3-5 years old, kept on commercial rations in a kennel. The drug was given to animals with food in a prophylactic dose (0.02 ml per 1 kg of animal weight) for 30 days. Blood samples were taken as animals for research before the use of the drug and at the end of the experiment. The intensity of the peroxidation processes was evaluated by the content of lipid peroxidation products in the blood (malondialdehyde - MDA, diene conjugates, dienketones). The activity of antioxidant enzymes, superoxide dismutase and catalase, was also determined. The research results are given in table.1-5.

Table 1 The effect of the use of the microelement preparation "Helavit" on the activity of dog blood catalase The indicator, unit rev. Before using chelavita After applying chelavita The number of red blood cells, 10 ¹² / l 5.14 ± 0.1 7.5 ± 0.5 Catalase number, mg H ₂ O ₂ 0.09 ± 0.05 0.13 ± 0.015 Catalase Index, Cat. number / qty er. in 1 μl of blood 0.017 ± 0.005 0.02 ± 0.005

The data in Table 1 show an increase in the number of red blood cells and catalase number, which indicates the normalization of erythropoiesis and an increase in the activity of the antioxidant enzyme.

table 2 Change in iron content and catalase activity Indicators, units rev. before application after application Iron, μmol / L 8.25 ± 1.06 14.16 ± 1.4 Catalase, units by bach 0.09 ± 0.05 0.13 ± 0.015 Table 3 Change in copper content and SOD activity Indicators, units rev. before application after application Copper, µmol / L 15.0 ± 0.04 37.5 ± 0.24 SOD, cu / mg protein per min 10.91 ± 0.63 12.8 ± 0.42 Table 4 Change in the concentration of lipid peroxidation products Indicators, units rev. before application after application MDA, μmol / L 22.2 ± 1.6 17.0 ± 1.0 Diene conjugates, units A / ml 0.25 ± 0.025 0.17 ± 0.07 Dienketones, units A / ml 0.15 ± 0.026 0.073 ± 0.005

From the data obtained it follows that when implementing the method, the concentration of copper and iron in the blood increases, which, in turn, leads to an increase in the activity of antioxidant enzymes such as catalase and superoxide dismutase. An increase in the activity of these enzymes is simultaneously accompanied by a decrease in the concentration of lipid peroxidation products, which indicates a decrease in the intensity of lipid peroxidation processes.

A change in the appearance of the animals was also observed. Before research, dogs had dull hair, and a number of animals had lower than average body weight. After the completion of the research, an improvement in the quality of the coat and an increase in body weight were observed.

Comparison of the effectiveness of the proposed method for the treatment of free radical pathology was carried out with the drug "Emitsidin" (a derivative of 3-hydroxypyridine and succinic acid). The use of emitsidin oral, large dogs 50 mg (2 capsules).

Table 5 Comparative efficacy of the use of emicidin and helavit preparations in the correction of oxidative stress (M ± m) Indicator Units measuring Before the use of drugs After applying chelavita After Emicidin SOD E. at./ mg protein per minute 10.91 ± 0.3 12.8 ± 0.4 11.3 ± 0.35 Catalase mg H ₂ About ₂ 0.09 ± 0.05 0.13 ± 0.015 0.11 ± 0.02 MDA μmol / l 22.2 ± 0.6 17.0 ± 0.45 18.23 ± 0.55 Diene conjugates units A / ml 0.15 ± 0.025 0.12 ± 0.07 0.13 ± 0.05 Dienketones units A / ml 0.25 ± 0.025 0.17 ± 0.07 0.18 ± 0.045

The data in table 5 indicate greater effectiveness of the proposed method compared with the use of the known antioxidant.

Example 2

The effectiveness of the claimed method was also carried out on horses aged 4-6 years. 2 groups of horses were selected, 5 animals each.

The drug "Helavit" was given to horses with food at a therapeutic dose of 0.06 ml per 1 kg of live weight 1 time per day for 30 days. In blood serum, the activity of catalase and SOD, as well as lipid peroxidation products, was determined analogously to example 1 (Tables 6, 7).

Table 6 Changes in the activity of SOD and catalase when using the microelement preparation Helavita horses Indicators, units rev. Before application After application Catalase, units by bach 7.64 ± 0.5 10.7 ± 0.75 SOD, cu / mg protein per min 8.25 ± 0.51 11.04 ± 0.94 Table 7 Changes in the concentration of lipid peroxidation products when using the microelement preparation Helavit in horses Indicators, units rev. Before application After application MDA, μmol / L 27.4 ± 1.7 22.4 ± 2.1 Diene conjugates, units A / ml 0.3 ± 0.04 0.21 ± 0.02 Dienketones, units A / ml 0.15 ± 0.9 0.09 ± 0.03

As can be seen from the data of Tables 6, 7, the implementation of the claimed method allowed to significantly increase the activity of enzymes responsible for antioxidant protection in the animal body, which accordingly reduced the concentration of lipid peroxidation products in the blood serum of test animals.

Thus, the data presented indicate the effectiveness of the proposed method for the treatment and prevention of oxidative stress in animals. The presence of a number of active trace elements in the chelated, bioavailable form in the preparation stimulated the synthesis of antioxidant enzymes such as superoxide dismutase, glutathione peroxidase, and catalase. Therefore, it is possible to use the proposed method for antioxidant therapy during pregnancy, aging, intensive growth and pathological conditions accompanied by oxidative stress in animals.

Claims (1)

A method of treating and preventing oxidative stress in animals, comprising administering to the animal a medicament containing selenium and active substances, characterized in that the microelement preparation Helavit is used as a medicament for prophylaxis in a daily dose of 0.01-0.02 ml per 1 kg of live weight of the animal orally with food with constant use, for treatment - in a daily dose of 0.05-0.07 ml per 1 kg of live weight for 4 weeks, followed by a transition to a prophylactic dose.