Method for increasing general nonspecific resistance of mammal body

FIELD: medicine.

SUBSTANCE: invention refers to experimental biology and medicine. The method involves parenteral introduction of a protein preparation. Ceruloplasmin is used as said protein preparation. Ceruloplasmin is introduced intravenously in dosage 7.5 mg/kg of animal body weight, for the total course of 3 injections every second day.

EFFECT: method allows intensifying anti-oxidant protection of an animal body, stimulating hemopoietic and immunological processes, and thereby providing aquatation of protective adaptive responses.

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The invention relates to the field of experimental biology and medicine and can be used to improve the overall nonspecific resistance of the organism mammals in various pathological conditions associated with the development of peroxidation-antioxidant imbalance and severe disorders of the immune status.

The known method of increasing the compensatory capacity of mammals, consisting in the inhalation of a hypoxic gas mixture, alternating with the inhalation of atmospheric air. Used part of the exhaled air to re-inhalation, reduce the consumption of hypoxic gas mixture prior to the onset of a mammal difficulty breathing. Then increase the hypoxic gas mixture to reduce shortness of breath and support the flow of hypoxic gas mixtures to levels that can produce moderate difficulty breathing mammal (RF patent No. 2108814, CL AM 16/00. Publ. 20.04.1998 year).

However, the known method is technically complex and time-consuming in execution.

There is also known a method of increasing the resistance of the organism in the experiment, namely, that hold the intake of 0.7-1.0 ml of blood from a vein, filgrastim dose of 10 mg/kg incubated with the blood 45 minutes at 37°C. followed by reinfusion (RF patent No. 2278690, CL AC 38/17, OR 37/02. Publ. 27.06.2006 g).

The disadvantage of this method is the complexity and difficulty of execution (the need for blood sampling, followed by incubation with the drug filgrastim and maintaining the resulting mixture at a specific temperature for a certain time) and intravenous reinfuse the mixture.

Closest to the proposed by its technical essence is a way to increase non-specific resistance, including intramuscular injection of nonspecific immunoglobulin with an additional appointment for the oral administration of biologically active stimulant in the form of an aqueous solution of 7-8%concentration at a dose of 2.0-2.5 ml/kg weight of the animal within 10-15 days with an interval of 24 hours (patent RF №2305551, CL AC 36/28, AK 36/53, AK 36/484, OR 37/00. Publ. 10.09.2007 BC).

The disadvantage of the prototype is the fact that for its implementation, in addition to intramuscular (parenteral) administration of the protein preparation is required for further use of a biologically active multi-component stimulator comprising extracts of herbs and roots. In addition, this method is time-consuming to conduct (10-15 days).

The challenge which seeks the invention is to simplify the method and shortening to achieve a positive result in increased non-specific resistant the spine of the body.

The problem is solved in that way increase the overall nonspecific resistance of the organism mammals by injecting the protein preparation, used as a protein drug use "Ceruloplasmin", administered intravenously at a dose of 7.5 mg/kg animal body weight, General course of 3 injections a day.

"Ceruloplasmin" ("Ceruloplasminum") - multifunctional medicinal drug, which is a glycoprotein alpha-2-globulin fraction of blood. The drug is approved for medical use in the Russian Federation (on the main page. Drugs in Russia: a Handbook. - M.: Attraversare, 2006, SB-1180).

Empirically, the authors claimed invention found that a dose of ceruloplasmin, a component of 7.5 mg/kg animal body weight is optimal for achieving the desired effect on the increase of resistance due to the improvement of peroxidation-antioxidant imbalance.

Dose of less than 7.5 mg/kg (5.0 mg/kg) does not provide adequate effect for improving the overall nonspecific resistance of the organism, and the dose of 7.5 mg/kg (10 mg/kg) does not give an additional positive effect to received.

The General course of treatment consisting of 3 intravenous injection, carried out through the day, to achieve optimal therapeutic effect and improve the state is of peroxidation-antioxidant imbalance in conditions of experimental arthritis in rabbits, confirmed by the data of instrumental and laboratory methods of research.

The technical result consists in the normalization of peroxidation-antioxidant balance by correcting the insolvency element of the antioxidant enzyme system.

The proposed method is as follows.

Rabbit with experimental rheumatoid arthritis is fixed on the machine for laboratory animals. Before drug injection is treated with 5%solution of iodine. Introduction ceruloplasmin is carried out in the marginal vein of the ear slowly sterile syringe at a dose of 7.5 mg/kg animal body weight, General course of 3 injections carried out through the day. After manipulation of the injection is sealed bactericidal plaster.

The inventive method was tested on 38 rabbits breed "Chinchilla Russian"was in standard vivarium conditions with free access to food and water. All experiments on animals were conducted in accordance with the standards of the Ethics Committee and the Declaration of Helsinki 1983

Experimental animals were divided into 5 groups:

1st group (control) - 7 intact (healthy) rabbits;

Group 2 (comparison) - 9 rabbits with experimental rheumatoid arthritis without treatment;

Group 3 (experienced) - 7 rabbits with experimental rheumato denim arthritis, who underwent a course of intravenous injection of ceruloplasmin in a dose of 5.0 mg/kg animal body weight, General course of 3 injections carried out in a day;

4-I group (experienced) - 8 rabbits with experimental rheumatoid arthritis who received a course of intravenous injection of ceruloplasmin in a dose of 7.5 mg/kg animal body weight, General course of 3 injections carried out in a day;

5th group (experienced) - 7 rabbits with experimental rheumatoid arthritis who received a course of intravenous injection of ceruloplasmin in a dose of 10.0 mg/kg animal body weight, General course of 3 injections carried out through the day.

Example. Rabbit No. 17 weight 3.4 kg was formed experimental rheumatoid arthritis according to the method Pettipher E.R. et al., (1987). The formation of the model was confirmed by clinical, instrumental and laboratory methods of research.

On visual inspection of the animal sedentary compared with intact rabbits. He had a reduced appetite, weight was 3,220 kg

The amount of experienced joint increased almost 1.5 times compared with symmetric joint. Due to swelling and severe pain reaction rabbit not relied on this limb.

Thermal analysis confirmed arthritis of the right knee joint. Detected temperature reaction in the area of the right knee joint was manifested by the difference in temperature is between ur experienced and symmetric joint and 0.9°C.

Along with the described clinical and imaging patterns diseases detected changes in laboratory parameters. The rabbit had metabolic disorders, both General and local character.

There is a strong overall activity of the inflammatory process, manifested by reduced hemoglobin - 118,20 g/l at norm 124,30±0,80 g/l; the number of red blood cells - to 3.33·1012/l at the rate of 4.04±0,10·1012/l; EMAS - 7,40 mm/h at the rate of 2.8±0.20 mm/h and the total number of leukocytes - 10,14·109/l at the rate of 7.60±0,11·109/l, and their band - of 5.82% at the rate of 3.96±0,18% and segmented - 51,37% at the rate 42,60±0,81% forms. Lymphocytes constituted of 33.26% at the rate 47,80±0,84%; eosinophils - to 2.57% at the rate 1,49±0,10%; monocytes - to 4.62% at the rate 3,44±0,12%.

Increased activity of enzymes (alanine aminotransferase - 29,4 E at the rate of 24.8 E; aspartate aminotransferase - 53,6 E at the rate of 48.5 E;

gamma-glutamyltranspeptidase - 5,7 E at the rate of 4.2 (E) testified to the development of liver damage, and increased total protein content to 61.8 g/l at norm - 56,35 g/l and urea to 7.02 mmol/l at norm - 5,95 mmol/l was speaking about violations of protein metabolism and renal function, respectively.

Also was discovered peroxidation-antioxidant imbalance lies in the activation of processes of lipid peroxidation (malonic dialdehyde - 5,2 MK is ol/l at norm 3,59±0.15 µmol/l) and insolvency enzyme level of the antioxidant system of blood (ceruloplasmin - 22,14 $ with the norm 27,77±1,03$).

The rabbit had a significant increase in the concentration of lactate in the blood - 17,12 mmol/l at norm 10,86±0.15 mmol/l and the pH decrease blood - 7,32 at the rate 7,42±0,01, testified to the disruption of the acid-base balance.

There was a statistically significant tension immunopathological reactions, defined by elevated levels of circulating immune complexes in serum - $ 21,10 compared with normal values for intact animals - 13,00±1,56 $

The total or aggregate severity of metabolic disorders were assessed in terms of molecules of average weight - 0,312 eop the norm 0,236±0,002 EDEP

Described denominated metabolic disorders identified in animals with experimental rheumatoid arthritis were similar with similar changes in patients with rheumatoid arthritis, which led to a reduction in the overall nonspecific resistance of the organism in this pathology.

This rabbit therapy using medicinal product "ceruloplasmin, enter in the vein slowly at a dose of 7.5 mg/kg animal body weight, General course of 3 injections a day.

After the treatment there was an improvement in the General condition of the animal. It was noted decrease swelling and temperature experienced joint. is in original condition. Palpation of the joint painless. The improvement of clinical picture of the disease was also confirmed by the data of laboratory tests.

There was a positive trend in the results of laboratory indicators: increase in the level of haemoglobin up of 122.5 g/l and the number of erythrocytes up to 3.58·1012/l, a reduction of ESR to 3.7 mm/h, total number of leukocytes to 8.3·109/l, stab neutrophils to 4.4% and segmented neutrophils to 46.1 per cent, and the reduction of eosinophils to 2,19%, monocytes before 3,81% and an increase of lymphocytes to 39,14%.

There was also a reduction in the activity of enzymes (alanine aminotransferase to 26.8 E, aspartate aminotransferase to 50.4 E, gamma-glutamyltranspeptidase to 4.8 (E), total protein content up to 59.0 g/l and urea to 6.84 mmol/l in the serum of the animal.

Indicators peroxidation-antioxidant balance was also positive: reduction of malondialdehyde to 4.7 mmol/l and increased activity of ceruloplasmin to 24.8 $

This rabbit has been an improvement in the blood pH to 7.35, reduction of circulating immune complexes to 16.00 USD, the level of lactate to 15,80 mmol/l and a content of molecules of average weight to 0,278 EDEP

Thus, the proposed method can improve the overall nonspecific resistance of the organism in conditions of experimental rheumatoid is rcrit, accompanied by the development of peroxidation-antioxidant imbalance and impaired immune status, which is possible due to intravenous injection of ceruloplasmin. This drug, showing antioxidant and immunomodulatory properties, launches molecular mechanisms that ensure the activation of the protective and adaptive reactions. The latter is achieved by resolving insolvency antioxidant protection system of the body, stimulation of hematopoietic and immunological processes, as well as decrease the severity of the intoxication of the organism as a whole.

The way to increase overall nonspecific resistance of the organism mammals by injecting the protein preparation, characterized in that is used as a protein drug use "Ceruloplasmin", administered intravenously at a dose of 7.5 mg/kg animal body weight, General course of 3 injections a day.



 

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