RU2762892C1 - Method for correcting microcirculatory disorders of the retina with asialated erythropoietin - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention relates to medicine, namely to experimental pharmacology and ophthalmology, and is intended for the correction of microcirculatory disorders in ischemic conditions of the retina. The method for correction of microcirculatory disorders of the retina in the experiment includes the introduction of asialated erythropoietin to a laboratory animal in an amount of 2.4 mcg/kg once on the 1st day of the experiment intraperitoneally 30 minutes before modeling the pathology.EFFECT: use of the invention improves the efficiency of correction of microcirculatory disorders in modeling retinal ischemia.1 cl, 1 tbl, 1 ex

Description

The invention relates to medicine, in particular to experimental pharmacology.

The closest to the claimed solution is a method for correcting retinal microcirculation disorders using recombinant erythropoietin "Method for the prevention of retinal ischemia with erythropoietin in experiment" (RU No. 2539629, publ. 01.21.2015), including modeling retinal pathology and prevention of it by introducing recombinant erythropoietin to a laboratory animal, the fact that the recombinant erythropoietin is administered as a preconditioning agent once on the 1st day of the experiment intraperitoneally at a dose of 50 IU / kg, 30 minutes before the modeling of the pathology.

The disadvantage of this method is that in many clinical situations requiring the cytoprotective effects of erythropoietin, its repeated administration is necessary, which leads to a significant increase in hematocrit, platelet activation and thrombus formation.

Accordingly, a promising direction is the use of substances capable of exerting non-erythrostimulating effects of recombinant erythropoietin and a preconditioning effect on the retina [Mennini T, De Paolo M, Bigini P, et al. Nonhematopoietic erythropoietin derivatives prevent motoneuron degeneration in vitro and in vivo. Mol Med 2006; 12: 153; Wang X, Zhu C, Wang X, et al. The nonerythropoietic asialoerythropoietin protects against neonatal hypoxia-ischemia as potently as erythropoietin. J Neurochem 2004; 91: 900.].

One of these substances is asialylated erythropoietin, a derivative of recombinant erythropoietin. Due to the complete absence of sialic acids in asialated erythropoietin, there is a short-term binding of this molecule to the heterodimeric erythropoietin receptor and the implementation of cytoprotective (non-erythropoietin) effects.

The objective of the present invention is a method for effective correction of retinal ischemia with asialated erythropoietin in an experiment, including asialated erythropoietin as a preconditioning agent.

The task is achieved by the fact that in a method including modeling retinal pathology by applying mechanical pressure (110 mm Hg) on the anterior chamber of the eye for 30 minutes and preventing it by administering asialated erythropoietin to a laboratory animal, and asialated erythropoietin is administered as a preconditioning agent once on the 1st day of the experiment intraperitoneally at a dosage of 2.4 μg / kg 30 minutes before modeling the pathology.

THE METHOD IS IMPLEMENTED AS FOLLOWS

The experiments were carried out on 40 white Wistar rats, sexually mature males, weighing 210-250 g. The experimental animals were divided into the following groups, each group included 10 rats:

Group I - a group of intact animals;

Group II - with retinal ischemia (control);

Group III - with the introduction of asialated erythropoietin at a dosage of 2.4 μg / kg;

Group IV - with the introduction of recombinant erythropoietin at a dosage of 50 IU / kg (reference drug).

Modeling of retinal ischemia was carried out by applying mechanical pressure (110 mm Hg) to the anterior chamber of the eye for 30 minutes on the 1st day of the experiment.

Asialated erythropoietin (Proteinovy Kontur LLC) was injected at a dosage of 2.4 μg / kg once intraperitoneally 30 minutes before modeling retinal ischemia. Recombinant erythropoietin (“Epocrine” epoetin alfa; Federal State Unitary Enterprise “State Research Institute of Highly Pure Preparations” of the Federal Medical and Biological Agency, St. Petersburg, Russia) was injected at a dose of 50 IU / kg once intraperitoneally 30 minutes before modeling retinal ischemia. 15 min before the modeling of ischemia, anesthesia was carried out by intraperitoneal injection of a solution of chloral hydrate at a dose of 300 mg / kg of the animal's body weight.

The severity of the correction of microcirculatory disorders of the retina was judged by the level of microcirculation using a Biopac-systems MP-150 laser Doppler flowmeter and a TSD-144 needle-type sensor (USA) at 72 hours of reperfusion after pathology modeling. The registration of LDF results was carried out by the Acqknowledge program version 3.8.1, the microcirculation values were expressed in perfusion units (PU).

Descriptive statistics were applied to all data. The data were checked for normal distribution. The distribution type was determined by the Shapiro-Wilk test. In the case of a normal distribution, the mean (M) and standard error of the mean (m) were calculated. Intergroup differences were analyzed using Student's t-test. Differences are considered significant at p <0.05. Calculations were performed using statistical programs Microsoft Excel 7.0.

EXAMPLE OF A SPECIFIC PERFORMANCE

Microcirculation was assessed using a Biopac-systems MP-150 laser Doppler flowmeter and a TSD-144 sensor (USA). For this, under anesthesia (chloral hydrate 300 mg / kg body weight, intraperitoneal), the animal was fixed and the level of microcirculation in the retina was recorded at ten points along the scleral circumference with a step of 1 mm. Recording of the curve of the level of microcirculation was carried out for 20 seconds at each point. From the ten values obtained, the mean was deduced, which was entered into the protocol and taken as the level of microcirculation in the retina in this animal. From 10 obtained values, the mean was deduced, which was taken as the level of microcirculation in the retina in this group of animals.

The results of assessing the level of microcirculation in the rat retina at 72 hours of reperfusion after modeling retinal ischemia and its correction with asialated erythropoietin at a dose of 2.4 μg / kg and recombinant erythropoietin at a dose of 50 IU / kg are presented in Table 1.

Table 1

The level of microcirculation in the retina of intact rats was 761.3 ± 9.6 p.u. (perfusion units). The level of microcirculation after modeling ischemia in the control group was 344.6 ± 10.2 p.u. at 72 hours of reperfusion, which is significantly lower than the value in the group of intact animals (p <0.001).

Against the background of pathology correction with asialated erythropoietin at a dose of 2.4 μg / kg, the level of microcirculation in the retina at 72 hours of reperfusion significantly increases to 746.8 ± 9.3 p.u. (p <0.001) compared with the control group. At the same time, the values achieved during the correction of pathology with asialated erythropoietin approach the values in the group of intact animals.

When the pathology is corrected with erythropoietin at a dose of 50 IU / kg, the level of microcirculation in the group increases to 710.7 ± 8.6 p.u. and significantly differs from the values in the control group (p <0.001).

Thus, the results obtained indicate a pronounced correction of microcirculatory disorders of the eye with asialated erythropoietin at a dosage of 2.4 μg / kg, under the conditions of the rat retinal ischemia model, with a single systemic administration 30 minutes before the pathology modeling.

Claims (1)

A method for correcting microcirculatory disorders of the retina with asialized erythropoietin in an experiment, including modeling of retinal pathology and correction of microcirculatory disorders of the retina by injecting erythropoietin into a laboratory animal once intraperitoneally 30 minutes before modeling the pathology, characterized in that asialated erythropoietin is used as erythropoietin. the amount of 2.4 μg / kg.