RU2372078C1 - Method for correction of endothelial dysfunction with preparation phosphogliv lyophilizate in l-name induced nitrogen oxide deficiency - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to experimental cardiopharmacology and can be used for correction of endothelial dysfunction. It is ensured by simulation of endothelial dysfunction in experiment by intraperitoneal introduction to Wistar rats of N-nitro-L-arginine methyl ester dosed 25 mg/kg within 7 days. Thus the degree of dysfunction development is estimated by the relation of endothelium independent and endothelium dependent vasodilatation. Dysfunction is corrected by intraperitoneal introduction of preparation phosphogliv lyophilisate dosed 5.0 ml/kg and 0.5 ml/kg.

EFFECT: invention provides effective correction of endothelial dysfunction in specific experimental conditions.

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Description

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

Closest to the claimed solution is a method of restoring endothelium-dependent cholinergic relaxation of the thoracic aorta of spontaneously hypertensive rats using phospholipid vesicles (liposomes), described by AISoloviev, AVStefanov, OVBazilyuk, VFSagach in the article "Phospholipid vesicles (liposomeerich dependent) relaxation in thoracic aorta from spontaneously hypertensive rats ”in J Hypertens. 1993, Jun; 11 (6): 623-7, which consists in the fact that phospholipid liposomes at a concentration of 100-125 μg / ml restore endothelium-dependent cholinergic relaxation of the thoracic aorta of spontaneously hypertensive rats, normalize the synthesis and secretion of the endothelial relaxing factor, and contribute to the correction of the endothelial function of damaged endothelial cells .

The main disadvantage of this method is the lack of bioavailability of a suspension of phospholipid liposomes and the risk of their absorption by the cells of the reticuloendothelial system, which reduce the ability of liposomes to reach certain target tissues. Therefore, in order to activate the correction of endothelial dysfunction, it is advisable to use the drug phosphogliv lyophilisate based on phospholipid and glycyrate, which restores the integrity and function of damaged endothelial cells. The optimal solution for the preparation of phosphogliv lyophilisate was the use of glycyrate not only as a pharmaceutical substance, but also as a detergent that promotes the production of stable nanoparticles that can integrate into the membranes of endothelial cells and effectively restore the activity of membrane enzyme systems and endothelial function.

The technical result of the invention is a method for correcting endothelial dysfunction, including the use of the drug phosphogliv lyophilisate. Before administration to the animals, the contents of one vial (2.5 g of lyophilisate, including phospholipid in an amount of 0.5 g and glycyrate in an amount of 0.2 g) were dissolved in 10 ml of water for injection. When calculating the doses, we used the coefficient of interspecific conversion of doses from a person to an experimental animal taking into account body weight (Ulanova I.P. et al., 1968).

The technical result is achieved by the fact that against the background of modeling endothelial dysfunction in an experiment by intraperitoneal administration to a laboratory animal for 7 days, a blocker of the synthesis of NO N-nitro-arginine-methol ether (L-NAME) at a dose of 25 mg / kg, its correction is carried out by intraperitoneal administration Phosphogliv lyophilisate at a dose of 5.0 ml / kg and 0.5 ml / kg (to prepare a solution, the contents of one bottle, i.e. 2.5 g of lyophilisate, are dissolved in 10 ml of water for injection) once a day.

The method is carried out as follows.

The experiments were carried out on white rats male Wistar line weighing 200-250 g. N-nitro-L-arginine methyl ether (L-NAME) was administered intraperitoneally at a dose of 25 mg / kg / day. On the 7th day from the start of the experiment, under anesthesia (ethinal sodium 50 mg / kg), a catheter is inserted into the left carotid artery to record blood pressure (BP), bolus administration of pharmacological agents is carried out in the right femoral vein. Hemodynamic parameters: systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) are measured continuously using the sensor and the Bioshell computer program. Functional tests: endothelium-dependent vasodilation (ESV) - intravenous administration of acetylcholine (AH) at a dose of 40 μg / kg, endothelium-independent vasodilation (ENZV) - intravenous administration of sodium nitroprusside (NP) at a dose of 30 μg / kg.

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

An example of a specific implementation.

A bolus intravenous administration of AH for 3-5 seconds led to a sharp drop in blood pressure, reaching a peak in intact animals for systolic pressure (SBP) 84.3 ± 4.4, for diastolic pressure (DBP) - 38.7 ± 2.8 and for mean arterial pressure (SRAD) 53.9 ± 2.7 mm RT. Art., during the first 2-3 seconds, a sharp bradycardia developed up to 130-150 beats per minute. Recovery of blood pressure occurred on average for 42.2 ± 0.8 seconds after normalization of the heart rhythm. ENZV was also characterized by a decrease in SBP to 83.0 ± 3.7, DBP to 42.1 ± 4.4 and SRAD to 55.7 ± 3.5 mm Hg. followed by complete recovery on average within 45.1 ± 1.0 sec. Blockade of NO synthase with the help of a long daily intraperitoneal injection of L-NAME (N-nitro-L-arginine methyl ester at a dose of 25 mg / kg) for 7 days caused arterial hypertension (SBP - 190.3 ± 6.7, DBP - 145.0 ± 3.9, SAD - 160.1 ± 4.6 mm Hg) and led to a smaller decrease in blood pressure after administration of AH (SAD to 110.6 ± 5.2, DBP to 82.8 ± 6.6, the control system up to 92.1 ± 6.1 mm Hg) and NP (the control system up to 88.7 ± 4.7, the DBP up to 50.8 ± 4.2 and the control system up to 63.4 ± 4 , 1 mmHg) compared with intact animals.

The simultaneous administration of L-NAME and phosphogliv lyophilisate at a dose of 5.0 ml / kg and 0.5 ml / kg (to prepare a solution of 2.5 g of lyophilizate was dissolved in 10 ml of water for injection) did not intraperitoneally decrease the initial BP values and amounted, respectively (SBP - 181.0 ± 10.5 and 203.2 ± 11.3, DBP - 138.3 ± 7.8 and 157.4 ± 8.4 mm Hg) compared with the untreated group animals (table 1.).

Table 1.Dynamics of blood pressure and heart rate in modeling nitric oxide deficiency and correction of endothelial dysfunction with phosphogliv (lyophilisate) based on phospholipid and glycyrate. Groups of animals Functional Tests GARDEN, mmHg DBP, mmHg Heart rate, beats in min Intact Source 137.7 ± 3.7 ^y 101.9 ± 4.3 ^y 420.0 ± 9.0 EZV with AH 84.3 ± 4.5 ^y 38.7 ± 2.8 ^y 416.0 ± 14.0 ENZV with NP 83.0 ± 3.7 42.1 ± 4.4 415.0 ± 10.0 Receiving L-NAME (25 mg / kg) Source 190.3 ± 6.7 ^* 145.0 ± 3.9 ^* 428.0 ± 11.0 EZV with AH 110.6 ± 5.2 ^* 82.8 ± 6.6 ^* 426.0 ± 14.0 ENZV with NP 88.7 ± 4.7 50.8 ± 4.2 426.0 ± 13.0 L-NAME (25 mg / kg) + phosphogliv lyophilisate (5.0 ml / kg) Source 181.0 ± 10.5 ^* 138.3 ± 7.8 * 364.6 ± 9.6 ^{* y} EZV with AH 95.0 ± 3.2 58.9 ± 4.0 ^{* y} 360.0 ± 9.3 ^{* Y} ENZV with NP 94.6 ± 4.9 47.8 ± 4.0 375.1 ± 10.2 ^y L-NAME (25 mg / kg) + phosphogliv lyophilisate (0.5 ml / kg) Source 203.2 ± 11.3 ^* 157.4 ± 8.4 * 391.3 ± 4.9 ^y EZV with AH 108.0 ± 5.1 65.1 ± 3.8 * 378.1 ± 4.2U ENZV with NP 120.7 ± 4.2 ^{* Y} 67.8 ± 7.8 392.5 ± 8.4 * - p <0.05 in comparison with the group of intact ^y - p <0.05 compared with the L-NAME group

The fundamental difference in EDV and ENZV in intact animals and animals that received an intraperitoneal inhibitor of NO synthase, L-NAME naturally led to the need to introduce a special indicator, the coefficient of endothelial dysfunction (QED), which characterizes the degree of endothelial dysfunction and is the ratio of the area of the triangle over the trend of the recovery reaction HELL in response to the introduction of NP to the area of the triangle above the trend of the reaction of HELL restoration in response to the administration of AH.

This ratio was calculated in each animal of the intact group and animals after modeling the blockade of NO synthase and obtained a difference of this indicator by 5 times — 1.1 in intact and 5.4 in animals treated with L-NAME, respectively.

Thus, to further evaluate the effect of AH and NP in ESV and ENZV, this ratio was used as an indicator of the correction of endothelial dysfunction. So, in groups that, against the background of the introduction of an inhibitor of NO-synthase L-NAME, received the drug phosphogliv lyophilisate at a dose of 5.0 ml / kg and 0.5 ml / kg (for the preparation of the solution, the contents of one bottle, i.e. 2.5 g of lyophilisate, dissolved in 10 ml of water for injection), QED was equal to 3.1 ± 0.4 and 2.9 ± 0.5, respectively lower than this indicator in the group of untreated animals (table 2).

Table 2.
Indicators reflecting the correction of endothelial dysfunction with the introduction of L-NAME, phosphogliv lyophilisate based on phospholipid and glycyrate. Groups of animals Functional Tests The increase in the fall of the vascular reaction by SRAD (mm Hg) Vascular reaction time (sec) The area of the vascular reaction (conventional units) The ratio of the areas of the vascular reaction of NP to AC Intact OH 59.9 ± 2.9 42.2 ± 0.8 ^y 1268.0 ± 74.8 ^y 1.1 ± 0.1 ^y NP 61.0 ± 3.0 ^y 45.1 ± 1.0 ^y 1375.3 ± 93.7 ^y L-NAME OH 68.0 ± 4.1 20.0 ± 1.4 ^* 695.3 ± 87.6 ^* 5.4 ± 0.6 ^* NP 98.0 ± 2.0 ^* 67.4 ± 1.4 ^* 3322.7 ± 116.7 ^* L-NAME + phosphogliv lyophilisate (5.0 ml / kg) OH 81.6 ± 6.1 ^* 42.0 ± 4.1 ^y 1710.8 ± 231.7 ^U 3.1 ± 0.4 ^{* Y} NP 97.0 ± 7.0 ^* 101.8 ± 6.8 ^{* Y} 4975.5 ± 578.8 ^{* y} L-NAME + phosphogliv lyophilisate (0.5 ml / kg) OH 93.3 ± 7.2 ^{* y} 49.0 ± 5.7 ^y 2347.0 ± 352.7 ^{* y} 2.9 ± 0.5 ^{* y} NP 86.0 ± 6.0 ^* 136.6 ± 12.3 ^{* y} 5783.7 ± 560.0 ^{* y} * - p <0.05 in comparison with the group of intact ^y - p <0.05 compared with the L-NAME group

Thus, the drug phosphogliv lyophilisate at a dose of 5.0 ml / kg and 0.5 ml / kg (for the preparation of the solution, the contents of one bottle, i.e. 2.5 g of lyophilisate, are dissolved in 10 ml of water for injection) when administered intraperitoneally once a day prevents the development of endothelial dysfunction with L-NAME-induced nitric oxide deficiency in the experiment.

Claims (1)

A method for the correction of endothelial dysfunction, characterized in that in an experiment with Wistar rats with endothelial dysfunction, against the background of its simulation by intraperitoneal administration of N-nitro-L-arginine methyl ester at a dose of 25 mg / kg daily for 7 days, assessing the degree of development of dysfunction by the ratio of endothelium-independent and endothelium-dependent vasodilation, daily phosphogliv lyophilisate is administered intraperitoneally at a dose of 5.0 ml / kg and 0.5 ml / kg.