RU2709833C1 - Method of placental microcirculation correction by asialized erythropoietin in adma-like model of preeclampsia - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely experimental pharmacology, and can be used for correction of placental microcirculation in ADMA-like model of preeclampsia. That is ensured by reproducing a model of preeclampsia in Wistar rats by daily intraperitoneal introduction of L-nitro-arginine-methyl ester 25 mg/kg from 14th to 20th day of pregnancy. That is combined with administration of asialized erythropoietin subcutaneously in dose of 2.4 mcg/kg once day from 10th to 20th day of pregnancy to correct the simulated pathology.

EFFECT: method provides pronounced correction of microcirculation in placenta in simulated pathology due to pronounced anti-ischemic and antioxidant action of administered asialized erythropoietin.

1 cl, 1 tbl, 1 ex

Description

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

Closest to the claimed solution is a method for correcting microcirculation disorders in the placenta using recombinant erythropoietin (RU No. 2466462, publ. 10.11.2012), which includes reproducing a gestosis model in Wistar rats by intraperitoneal administration of L-nitro-arginine daily from 14 to 20 days of gestation methyl ester in a dose of 25 mg / kg, and for the correction of the simulated pathology, recombinant erythropoietin is administered subcutaneously at a dose of 50 IU / kg on the 7th, 10th, 13th, 16th, 19th day of pregnancy.

The disadvantage of this method is that by binding to the homodimeric receptor, erythropoietin can activate erythropoiesis, which leads to a violation of the rheological properties of the blood and is an undesirable phenomenon in ischemia (Enhancement of ventricular-subventricular zone-derived neurogenesis and oligodendrogenesis by erythropoietin and its derivatives / N. Kanek , E. Kako, K. Sawamoto // Front Cell Neurosci. - 2013 .; Erythropoietin: recent developments in the treatment of spinal cord injury / S. Carelli, G. Marfia, A. Maria Di Giulio).

This fact makes it impossible to increase the dose of erythropoietin with its low efficiency. Therefore, the results of microcirculation correction in an ADMA-like model of pathology in pregnant animals using recombinant erythropoietin are unsatisfactory.

One of the ways to eliminate this drawback is the creation of erythropoietin derivatives with a short period of life. To activate the cytoprotective effects of erythropoietin, it is sufficient to contact the heterodimeric receptor for a short time (5 minutes). To activate erythropoiesis, a long-term continuous presence of erythropoietin at the homodimeric receptor is required (Enhancement of ventricular-subventricular zone-derived neurogenesis and oligodendrogenesis by erythropoietin and its derivatives [Text] / N. Kaneko, E. Kako, K. Sawamoto // Front Cell Neurosci - 2013; Erythropoietin: recent developments in the treatment of spinal cord injury [Text] / S. Carelli, G. Marfia, A. Maria Di Giulio [et al.] // Neurol Res Int. - 2011; The use of erythropoietin and its derivatives to treat spinal cord injury [Text] / A. Mofidi, A. Bader, S. Pavlica [et al.] // Mini Rev Med Chem. - 2011).

However, despite the obvious prerequisites, the question of the effectiveness of the derivative under study in connection with the change in its lifetime remains ambiguous. This is due to the fact that in various tissues and organs metabolic, regulatory and other processes have their own not only temporary, but also other features. The placenta is a dynamically developing organ with a complex combination of organogenesis and apoptosis. In addition, changes in the spatial structure, mass and charge also leave open the question of its permeability through tissue barriers (and in particular fetoplacental).

In connection with the foregoing, the object of the invention is to provide a more effective method for correcting microcirculation disorders with an ADMA-like model of preeclampsia, including the use of asialized erythropoietin (Protein circuit LLC), which has a short half-life.

The problem is solved using the proposed method in which, against the background of simulating preeclampsia in an experiment, intraperitoneal administration to pregnant female Wistar rats for 7 days of an ADMA-like blocker of endothelial NO synthase - N-nitro-L-arginine-methyl ether (L-NAME) at a dose of 25 mg / kg, microcirculation disorders are corrected by subcutaneous administration of asialized erythropoietin at a dose of 2.4 μg / kg 1 time per day from 10 to 20 days of pregnancy. This leads to a pronounced correction of microcirculation in the placenta with simulated pathology. This effect is associated with a pronounced anti-ischemic and antioxidant effect of asialized erythropoietin.

The method is carried out as follows.

The experiments were carried out on white pregnant rats of Wistar females weighing 250-300 g. N-nitro-L-arginine methyl ether (L-NAME) was administered intraperitoneally at a dose of 25 mg / kg / day for 7 days (from 14 to 20 days of pregnancy ) Asialized erythropoietin is administered subcutaneously from 10 to 20 days of pregnancy at a dose of 2.4 μg / kg 1 r / day.

On the 21st day of pregnancy under anesthesia, microcirculation in the placenta was measured. The placenta microcirculation was studied using Biopac systems equipment: MP100 polygraph with laser Doppler flowmetry (LDF) LDf100C module and TSD144 sensor. The LDF results were recorded using the Acqknowledge version 3.8.1 program; microcirculation values were expressed in perfusion units (PED).

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

EXAMPLE OF SPECIFIC IMPLEMENTATION.

Blockade of NO synthase caused by 7-day administration of L-NAME to pregnant rats led to impaired microcirculation in the placenta, as evidenced by a decrease in its value from 492.8 ± 21.07 PED in intact pregnant animals to 211.8 ± 6.03 PED (p <0.05) in the control group. Subcutaneous administration of asialized erythropoietin at a dose of 2.4 μg / kg 1 r / day from 10 to 20 days of pregnancy against the background of an ADMA-like model of preeclampsia in pregnant rats also led to a significant increase in the level of microcirculation in the placenta to 443.2 ± 22.96 PED, which is higher than in animals in the group with the introduction of L-NAME (p <0.05), and slightly lower than in animals in the intact group.

Thus, the obtained results convincingly indicate a pronounced correction of the microcirculation disturbance in the placenta under the conditions of the similar model of pre-eclampsia in pregnant rats reproduced by ADMA with asialized erythropoietin. The effect of asialized erythropoietin on blood pressure and microcirculation are shown in table 1.

Table 1.

Claims (1)

A method for correcting microcirculation disorders in the placenta with an ADMA-like model of pre-eclampsia, including reproducing the pre-eclampsia model in Wistar rats daily from 14 to 20 days of pregnancy by intraperitoneal administration of L-nitro-arginine-methyl ether at a dose of 25 mg / kg, characterized in that for correction of simulated pathology is administered asialized erythropoietin subcutaneously at a dose of 2.4 μg / kg 1 time per day from 10 to 20 days of pregnancy.