Method for modeling hypoxic encephalopathy

FIELD: experimental medicine.

SUBSTANCE: laboratory animals should be once injected intraperitoneally or intravenously with phenylhydrazine at the dosage of 100-150 mg/kg.

EFFECT: higher efficiency.

1 cl, 4 ex, 2 tbl

The invention relates to medicine, specifically to experimental medicine, and relates to a method of modeling hypoxic encephalopathy.

Known methods of modeling hypoxic encephalopathy, by ligating the brain vessels, a temporary cardiac arrest, or placement of animals in thermocamera [1, 2], the last is the closest to the positive effect of the method of modelling of hypoxic encephalopathy prototype.

However, these methods are very time consuming, not reproducible and are often not adequate to study the pathogenesis of diseases associated with hypoxia.

The problem solved by this invention is the simplification of the method, the higher degree of reproducibility and the extension of the scope of the model of hypoxic encephalopathy.

This object is achieved in that laboratory animals (mice or rats) once intraperitoneally or intravenously administered at the maximum tolerated dose of hemolytic poison, which is used as the phenylhydrazine at a dose of 100-150 mg/kg

New in the present invention is that to simulate hypoxic encephalopathy using phenylhydrazine at a dose of 100-150 mg/kg

The authors are not found in the analyzed literature, fashion modeling hypoxic encephalopathy by Hemi is eskay hypoxia.

Comparison of the invention with existing methods of modeling encephalopathy showed that for the first time proposed to simulate hypoxic encephalopathy by administering to animals the maximum tolerated dose hemolytic venom, which is used as the phenylhydrazine at a dose of 100-150 mg/kg

Factor in the possibility of obtaining the desired result by administering to animals the maximum tolerated dose hemolytic poison for the specialist is not obvious. The experiment showed unexpected results.

Thus, the claimed invention meets the criteria of the invention of “Novelty” and “Inventive step”because it is not obvious to a person skilled in the art. The present invention meets the criterion of “Industrial applicability”because it can be successfully used in experimental medicine (physiology, pathological physiology, pharmacology) to simulate pathological conditions in humans and animals.

Hypoxia, disrupting energy metabolism, causes damage to, first of all structures of the Central nervous system, identified specific metabolism and neuronal functioning. The main feature of the latter is the combination of high metabolic activity, coupled with intensive consumption to the of Sloboda and high update rate Fund high energy substances. Inhibition of aerobic oxidation processes in the brain during hypoxia significantly reorganizes the Central nervous system, which creates prerequisites for integrative changes-starting activity of neurons and after exposure, and in the case of decompensation of adaptation mechanisms, triggering a chain of pathological processes leading to progressive changes in metabolic and functional performance of many internal organs. On the other hand, in the post-hypoxic period, the brain itself becomes the object of the pathogenic effects of organ pathology: systemic hemodynamic changes, disorders of the immune status, blood system, hemostasis, etc. that contributes to the aggravation of the damage to the Central nervous system.

The method is as follows:

The animal once intraperitoneally or intravenously administered at the maximum tolerated dose of hemolytic poison, which is used as the phenylhydrazine at a dose of 100-150 mg/kg

The proposed method was studied in experiments on mice CBA/CaLac (animals of category 1, the conventional linear mouse) in an amount of 80 pieces, weighing 18-20 g, and 18 outbred rats. Animals obtained from the nursery of the Department of experimental biomedical modeling, Institute of pharmacology, Siberian branch of the Russian Academy of medical Sciences (certificate available).

Example 1.

Sososososososo as follows. The mice CBA/ CaLac, weighing 18-20 g, once intraperitoneally injected with a phenylhydrazine in a dose of 150 mg/kg

Example 2.

The mice CBA/ CaLac, weighing 18-20 g, once intravenously injected with a phenylhydrazine in a dose of 100 mg/kg

Example 3.

Outbred rats, weighing 250-300 g, once intraperitoneally injected with a phenylhydrazine in a dose of 150 mg/kg

Example 4.

Outbred rats, weighing 250-300 g, once intravenously injected with a phenylhydrazine in a dose of 100 mg/kg

Hypoxic exposure was modeled by a single intravenous injection of phenylhydrazine at a dose of 100-150 mg/kg

Assessment of the condition of the Central nervous system of animals produced by recording the neuropsychiatric status: conditional reflex activity and orienting-exploratory behavior of animals in the open field.

After 1 day after injection of phenylhydrazine in all cases, animals have developed a conditioned reflex of passive avoidance [3], checking safety reflex was performed on the 7th, 14th, 21st day of the experiment. Check indicators orienting-exploratory behavior in the open field was made on the 3rd, 7th, 14th and 21st day after exposure [4].

Analysis was performed by the method of variation statistics using t-test, t-test and non-parametric U-test, Wilcoxon-Mann is Whitney [5].

The results of the study.

During the experiment it was shown that intraperitoneal or intravenous injection of phenylhydrazine at a dose of 100-150 mg/kg resulted in significant changes in locomotor activity of animals in the open field and the development of obvious violations of mnestic functions of the Central nervous system of mice.

Thus, the study tentatively research behavior after exposure showed an increase of the asymmetry factor movements on 3, 7, 14, 21 St day of research, by increasing the amount of horizontal movement of animals on the 7th, 21st day, and the fall in the number of surveys holes of the field on the 14th day of the experience, as well as reducing the number of acts of grooming on the 3rd day (table. 1).

In addition, there was a statistically significant decrease in the playback level of the conditioned reflex of passive avoidance on the 7th, 14th, 21st day of the study (up to 0% on the 21 St day) and spontaneous death of animals up to 20% on the 7th day of the experiment (table. 2).

Thus, intraperitoneal or intravenous injection of phenylhydrazine at a dose of 100-150 mg/kg resulted in signs of pathology of the Central nervous system characteristic of encephalopathy.

The proposed method allows to simulate hypoxic encephalopathy by introducing experimental animals at the maximum tolerated dose of hemolytic poison, as is e which is used as a phenylhydrazine at a dose of 100-150 mg/kg, that allowed us to simplify the model, to improve reproducibility and to expand the scope of application, in particular, to use in Hematology research.

Literature

1. Methodical recommendations on scringe and preclinical testing antihypoxic funds./ Ugoku Associ etc., M., 1989. - 20 S.

2. Suslov NI Pathogenetic substantiation of the psychopharmacological effects of drugs of natural origin: Dis... Prof. the honey. Sciences. - Tomsk, 1995. - 406 S.

3. Bureš J., Buresova O., Houston J. Pettite and basic experiments for the study of brain and behavior. / Lane. from English. Edited by Prof. Assatova). - M.: Higher school, 1991. - 398 S.

4. Walsh R.N., Cummins R.A. The open-field test: a critical review. // Psychol. Bull. - 1976, V.83. - P. 482-504.

5. Lakin GF Biometrics. - M.: Higher school, 1973. - S.

1. The simulation method hypoxic encephalopathy, consisting in the introduction of experimental animals of phenylhydrazine at a dose of 100-150 mg/kg

2. The method according to claim 1, characterized in that the phenylhydrazine injected once intraperitoneally or intravenously.