Method of prevention of acute radiation sickness in animals

 

(57) Abstract:

The method applies to radiobiology and veterinary biotechnology and is intended for the prevention of acute radiation sickness in animals. The essence of the method: in the introduction to the animals subcutaneously at a dose of 1.0 mg/kg twice for 30 and 14 days prior to acute radiation exposure of tissue preparation with a concentration of 10 mg/ml protein, obtained by the bleaching of the tissue homogenate of the liver, spleen, lymph nodes and blood serum of pigs, sheep and other animals killed after 3 to 7 days after their irradiation with the dose of radiation that causes acute radiation sickness is severe. 3 table.

The invention relates to radiobiology and veterinary biotechnology, in particular to a method for prevention of acute radiation sickness in animals, and can be used in the development of radioprotective activities in animal husbandry in relation to the conditions of mass exposure to ionizing radiation (accident, etc).

In the literature there are data on the use of biologically active substances from the tissues of animal origin with radioprotective properties.

Purified extract of spleen calves mol. weight 50 - 75 KD, survival of animals (Olontseva O. N. Purified splenic extract - treatment of radiation sickness in the experiment. //Radiation damage to the body and ways of their correction /Institute of biology and Biophysics, Tomsk state University, Tomsk, 1991, 28 - 31).

Immunocorrective preparation from bone marrow when introduced subcutaneously irradiated with gamma rays at doses of 6 and 6.5 Gy mice increased their survival, due to the ability of the drug to speed up the process of differentiation of hematopoietic precursors, immunocompetent cells in the bone marrow (immunocorrective Effect of the drug mielopid on some indicators of the blood and immune systems in experimental acute radiation sickness (Stepanenko R. N., Badinan S. A., frost, B. B. and others// Immunology, 1993, N 3, p. 25 - 28).

Has radioprotective effect of vilozena and thymogen (both drugs from the thymus of animals) under various schemes of their introduction (Baraboi C. A., yalqut S. I., Zorin centuries of low Molecular weight factors of thymus, as a means of prevention and early pathogenetic therapy of radiation injuries // Radiobiological Congress, Kiev, 25 Sept., 1993: Abstracts, so 2, Pushchino, 1993, S. 80 - 81). Double introduction of vilozena intraperitoneally after 2 and 24 h after UV irradiation is.

Known low-molecular substances, the weight of 10 KD (splenopathy) isolated from the spleen of pigs killed in 3 - 5 days after irradiation in doses 3,0-5,0 G (u C. C. Pathophysiological rationale some ways purposeful search pharmacological means of radiation protection: author. dis. Dr. Biol. Sciences. Obninsk, 1989). Splenopathy isolated from the spleen of irradiated pigs after subcutaneous administration to mice for 1 - 3 days before irradiation resulted in a significant increase in survival of animals compared to control exposure, which is associated with a favorable effect on the recovery of hemopoiesis.

Known methods of application and other radioprotective biologically active agents, each representing a different peptides and belonging to the class of active regulators of hemopoiesis.

However, the limited application of biological preparations in this class in a narrow time period relative to the time of irradiation significantly reduces the possibility of holding radioprotective activities.

The proposed method is the use of tissue preparation eliminates these disadvantages.

The objective of the invention is a method of preventing acute radiation sickness animals tnim subcutaneously at a dose of 1.0 mg/kg twice for 30 and 14 days prior to acute radiation exposure of tissue preparation with a concentration of 10 mg/ml protein, obtained clarification of the tissue homogenate of the liver, spleen, lymph nodes and serum of pigs, sheep and other animals killed after 3 to 7 days after irradiation in the dose that causes acute radiation sickness severe.

To obtain tissue preparation use of animals (pigs, sheep, and others ). Total uniform gamma-irradiation of animals produce gamma settings rays of Cs-137, once in doses of LD 50-100/30 (0,9 - 1,8 10-3CL/kg) at a dose rate of 0.9 to 1.4 R/min (3,8 - 6,0 10-6CL/(kgf) and the uniformity of the field of irradiation of 10%. The uniformity of the exposure to radiation is provided by moving the platform with the animals at a constant speed between sources.

After exposure for 3 to 5 days in animals deprived blood to obtain serum and after slaughter remove the liver, spleen and lymph nodes. Subsequent stages of work on the preparation of the complex radiation antigens is carried out with observance of the conditions of sterility.

Washed naliczenie organs from the blood with saline. The fabric selected organs scissors cut into small pieces, then crush using an electric grinder and homogenized with the addition of distilled water in sooie 30 minutes To the obtained supernatant bodies add the whey and determine in a mixture of the protein. Saline solution is brought to a final concentration of 10 mg/ml, poured into vials of 5 - 10 ml and frozen at -40oC. For longer storage material is subjected to freeze drying: freeze the material at -50oC for 3 h, maintained at a temperature of from -50oC and 0oC for 50 hours Then bring the temperature up to 201oC for 3 h and finally dried 3 hours Stored at 8 to 10oC.

The dried preparations are kind of dry porous mass of yellowish-gray color.

Sterility is defined at all stages of production of biological drug by planting on the BCH, MPA and environment Saburo.

The specificity and activity of preparations to ensure that calcarenitic, RDP and IAAF using sera obtained from pigs, gipomaniakalnah tissue radioprotective drug. As a control, use similar preparations, obtained from the tissues of intact animals.

The drug has a titer of 1:512 in the reaction of calcarenitic, and when determining specificity in RDP 1:8 with cross rustically with double breeding ant is ity of the drug, evaluated in the reaction of splenic colony, is not less than 1.8 (P<0,05).

The drug has a pronounced protective radioprotective action: the survival of irradiated animals is 100% at the end of the 60 - 67% only irradiated animals.

Example 1. The experiments were conducted on 38 outbred rats weighing 180 - 200 g Rats were divided into 4 groups (table. 1):

1st group - the drug was administered twice for 30 and 14 days before irradiation subcutaneously at 5 mg per rat;

2nd group - the drug was administered for 7 days before irradiation once subcutaneously with 5 mg per rat;

group 3 served as a control exposure;

4 - biological control (not irradiated).

Irradiation was carried out with gamma rays of cesium-137 at a dose of 700 R dose 1,41 R/min

Observation of the animals was carried out within 60 days.

After the injection the rats were not observed variance in the overall health, death is not recorded in the month. The use of the drug for 30 days with the re-introduction of 2 weeks was allowed to protect 100%, for 7 days - 75%, with a weight of 18 - 20 g were divided into 3 groups (table. 2):

1st group - the drug was administered twice (within 30 days before irradiation with subsequent introduction of a through 14 d) and 1 mg per mouse subcutaneously;

group 2 served as a control exposure;

group 3 - biological control (irradiation were not exposed).

Irradiation of animals was performed with gamma rays cesium-137 dose of 1.4 R/min at a dose of 700 R.

Observation of the animals was carried out within 60 days.

With the introduction of the drug to mice during the month were not observed deviations in the shared state, the death of the animals. Study drug was allowed to protect up to 100% of mice at the end of 66.6% of the animals in the control group exposure.

Example 3. The experiments were conducted on 6 pigs of large white breed weighing 20 to 25 kg, divided into 2 groups (table. 3):

1st group - the drug was administered twice for 30 and 14 days before irradiation subcutaneously at 1 mg/kg;

group 2 served as a control exposure.

Irradiation of animals was performed with gamma rays cesium-137 dose to 1.21 P/min at a dose of 400 R.

Observation of the animals was carried out for 75 days.

After the injection of the variance in the overall health of pigs is not marked. The drug helped to keep all StatLine on the course of radiation sickness clinical signs were less pronounced, to a lesser extent compared to control animals, a marked reduction in the number of leukocytes, lymphocytes and their populations.

The invention can be used for the prevention of acute radiation sickness in animals.

Method of prevention of acute radiation sickness in animals, including the introduction of the drug obtained from irradiated animals, characterized in that, for the prevention of acute radiation sickness, use of tissue radioprotective drug consisting of the clarified homogenate mixture of tissues of liver, spleen, lymph nodes and blood serum of irradiated animals with a protein concentration of 10 mg/ml, which is injected subcutaneously at a dose of 1.0 mg/kg twice for 30 and 14 days. to radiation exposure.

 

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