RU2466463C2 - Method for pharmacological correction of extremity ischemia - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: for the purpose of studying of the potential correction of ischemia, extremity ischemia is simulated in experimental animals. It is followed by the daily 28-day intragastric introduction of mixed solutions of homoeopathic dilutions of rabbit monoclonal antibodies to endothelial nitrogen oxide synthase -C12, C30, C200 4.5 ml/kg twice a day.

EFFECT: effective ischemia treatment ensured by stimulated neovasculogenesis, as well as eliminated side effects, as the applied pharmacological agent is related to a class of preparations of very-low-dose antibodies.

1 ex, 1 tbl

Description

The invention relates to medicine, in particular to experimental pharmacology, and can be used to correct limb ischemia.

Closest to the claimed solution is a method of correcting limb ischemia (RU 2302239, publ. 07/10/2007), which consists in the fact that in an experiment a laboratory animal with experimental limb ischemia is injected intraperitoneally with L-arginine once from the 7th to the 28th day after modeling ischemia in a dose of 30 to 200 mg / kg.

A disadvantage of the known solution is that the administration of L-arginine causes side effects, namely embryotoxicity, teratogenicity and mutagenicity.

The objective of the invention is to develop an effective method for the correction of limb ischemia, which eliminates the presence of side effects.

This object is achieved in that in an experiment for a laboratory animal with experimental limb ischemia, the correction of limb ischemia is carried out by administering a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial oxide synthase daily for 28 days after modeling ischemia, twice a day nitrogen - C12, C30, C200 at a dose of 4.5 ml / kg (9 ml / kg / day).

The main advantage of the proposed method is the absence of embryotoxicity, teratogenicity and mutagenicity of the pharmacological agent, since it belongs to a new class of ultra-low dose preparations (SMD) of antibodies.

Interdisciplinary studies conducted over a period of more than 6 years in a number of research and clinical institutions of Russia have revealed the main mechanisms underlying the biological and clinical effects of ultra-low doses (SMD) of antibodies administered orally. The mechanism of action of a mixture of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200 (the active substance in the preparation is diluted 100 ¹² , 100 ³⁰ and 100 ²⁰⁰ times, respectively) is their activation by the principle of "feedback" endothelial NO- synthases. Under the action of this enzyme, endothelial cells secrete nitric oxide II (NO), a powerful endogenous humoral vasodilator that is involved in vascular relaxation and stimulates the growth of new vessels (neovasculogenesis). Nitric oxide II diffuses into the smooth muscle cells of blood vessels, which leads to their relaxation. Preclinical studies have confirmed the absence of embryotoxicity, teratogenicity and mutagenicity in preparations of ultra-low doses of antibodies (Epstein, O. I. Pharmacology of ultra-low doses of antibodies to endogenous function regulators: Ph.D. Dr. O. Epstein. - Tomsk, 2003. - S.352).

The method is as follows.

The experiments were carried out on white Wistar rats weighing 250-300 g. Limb ischemia was simulated under anesthesia (chloral hydrate 300 mg / kg ip) by removing a section of the main vessel, including the femoral artery, popliteal artery, and the initial parts of the leg artery.

Microcirculation in the muscles of the tibia of an animal is assessed using a Biopac-systems MP-100 laser-Doppler flowmeter and TSD-144 sensor (USA). To do this, under anesthesia (chloral hydrate 300 mg / kg of weight intraperitoneally), a section of the skin in the anterolateral region of the leg region is excised. The microcirculation level is recorded at five points (the middle of the muscle length, points 3-5 mm above and below, lateral and medial of the first).

Registration of the level of microcirculation is carried out in groups of animals: intact; falsely operated; with modeling of limb ischemia (control); with correction of limb ischemia with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200.

A mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200 was administered intragastrically (via a probe) daily at a dose of 4.5 ml / kg 2 times a day for 28 days after modeling of ischemia.

When statistical data processing is calculated, the average value, the standard deviation. Differences are considered significant at p <0.05.

An example of a specific implementation.

The level of microcirculation in the right and left muscles of the leg of intact animals did not differ from each other and amounted to 525.7 ± 9.8 p.u. (perfusion units). Experimental animals were divided into the following groups:

1. intact (n = 10);

2. falsely operated (n = 10);

3. with modeling of ischemia of the leg muscle (n = 10);

4. with modeling of ischemia of the leg muscle + a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200 (n = 10).

Simulation of ischemia of the leg muscles in rats was carried out under anesthesia (chloral hydrate 300 mg / kg ip) by removing a section of the main vessel, including the femoral artery, popliteal artery, and the initial parts of the leg arteries.

Correction of limb ischemia was performed by intragastric (via a probe) injection of a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200 at a dose of 4.5 ml / kg twice a day (9 ml / kg / day) every day 1st to 28th day after modeling of ischemia.

Microcirculation in the muscles of the calf of the animal was assessed using a Biopac-systems MP-100 laser-Doppler flowmeter and a TSD-144 sensor (USA) on day 28. To do this, under anesthesia (chloral hydrate 300 mg / kg of weight intraperitoneally), a section of skin was excised in the anterolateral region of the leg. The microcirculation level is recorded at five points (the middle of the muscle length, points 3-5 mm above and below, lateral and medial of the first). The microcirculation level curve was recorded for 30 seconds at each point. From the five values obtained, the average was deduced, which was entered into the protocol and was taken as the level of microcirculation in the calf muscles in this animal. From 10 obtained values, the average was deduced, which was taken as the level of microcirculation in this group of animals at a given time of the study.

The results of assessing the level of microcirculation in animals on day 28 when modeling limb ischemia and correcting it with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200 at a dose of 9 ml / kg / day are presented in the table daily (table 1 )

Table 1 The results of assessing the level of microcirculation in the leg muscles on day 28 (M ± m), p.u. (perfusion units) Intact (n = 10) 525.7 ± 9.8 False operated (n = 10) 531.8 ± 12.8 Limb Ischemia (n = 10) 361.3 ± 8.7 * Limb ischemia + a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200 (n = 10) 490.3 ± 18.6 ** Note: * - p <0.05 in comparison with the intact group of animals; ** - p <0.05 compared with a group of animals with limb ischemia

The table shows that the level of microcirculation after the operation of modeling ischemia of the hind limb was 361.3 ± 8.7 p.u. by 28 days, which is significantly lower than the values in the group of intact (p <0.001) and falsely operated animals (p <0.001). Against the background of pathology correction with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200, the level of microcirculation significantly increases to 490.3 ± 18.6 p.u. (p <0.05).

Thus, the results obtained allow us to ascertain the effectiveness of the pharmacological correction of limb ischemia with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200 due to the stimulation of neovasculogenesis. In addition, the claimed method of pharmacological correction of limb ischemia allows to exclude side effects, since the pharmacological agent used belongs to the class of ultra-low doses of antibodies.

Claims (1)

A method for pharmacologically correcting limb ischemia, including administering to a laboratory animal a pharmacological agent after ischemia modeling, characterized in that a mixture of solutions of homeopathic dilution of polyclonal rabbit antibodies to endothelial nitric oxide synthase C12, C30, C200, which is intragastrically in a dose of 4, is used as such a drug 5 ml / kg (9 ml / kg / day) is administered to the laboratory animal daily, for 28 days after modeling of ischemia, twice a day.