RU2301670C1 - Method for correction of endothelial dysfunction by combination of enalapril and resveratrol in l-name-induced deficiency of nitrogen oxide - Google Patents

Abstract

FIELD: medicine, experimental cardiopharmacology.

SUBSTANCE: method involves modeling the endothelial dysfunction by every day intraperitoneal administration of L-nitroarginine methyl ester in rats (Wistar strain) in the dose 25 mg/kg for 7 days. On background of dysfunction modeling its correction is carried out by simultaneous intragastric administration of enalapril in the dose 0.5 mg/kg and by intraperitoneal administration of resveratrol in a single dose 2 mg/kg per 24 h. Degree of dysfunction development and activation of its correction are evaluated by the ratio endothelium-independent and endothelium-dependent vasodilation. Method provides possibility for study of endothelium-protective effects of resveratrol and activation of endothelial NO-synthase at stage of modeling endothelial dysfunction and their evaluation by the above given ratio endothelium-independent and endothelium-dependent vasodilation. Invention can be used in correction of endothelial dysfunction.

EFFECT: improved method of correction.

2 tbl, 1 ex

Description

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

Closest to the claimed solution is a method for the correction of endothelial dysfunction using enalapril, which belongs to ACE inhibitors, described by Ageev F.T., Ovchinnikov A.G. "Angiotensin-converting enzyme inhibitors in the treatment of cardiovascular disease. (Quinapril and endothelial dysfunction)," Chapter II. Endothelial Dysfunction, Part V. ACE inhibitors for endothelial dysfunction: experimental data. M., 2001. Pfizer Publishing House, http://www.viagra.nnov.ru/info/10911-ingibitors4.html |, which consists in the oral administration of these drugs in patients to correct endothelial dysfunction.

The main disadvantage of this method is the narrow pathogenetic orientation and mediation of enalapril during endothelial dysfunction, associated with its low affinity for the tissue (endothelial) reninangiotensinaldosterone system (RAAS), while the main reason for the development of this pathophysiological state is a decrease in the activity of vascular endothelial cells in the formation of nitric oxide (NO), remains uncompensated. Therefore, the results of the correction of endothelial dysfunction with a lack of NO synthases using enalapril are unsatisfactory.

The technical result of the invention is a method for the correction of endothelial dysfunction, including the use of enalapril, an ACE inhibitor, and an activator of NO synthase, resveratrol.

The technical result is achieved by the fact that, against the background of modeling endothelial dysfunction in an experiment, an intraperitoneal injection of a blocker of NO synthesis of L-nitro-arginine methyl ether (L-NAME) at a dose of 25 mg / kg is administered intraperitoneally to a laboratory animal for 7 days, and correction is carried out by intragastric administration of enalapril at a dose of 0.5 mg / kg once a day and intraperitoneal administration of the resveratrol NO synthase activator at a dose of 2 mg / kg also once a day, which leads to increased correction of endothelial dysfunction, since resveratrol is they are activated by the NO synthase enzyme by which NO is formed in the vascular endothelium.

THE METHOD IS CARRIED OUT AS FOLLOWS.

The experiments were carried out on white rats male Wistar line weighing 250-300 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, a catheter is inserted into the left carotid artery under anesthesia (etaminal sodium 50 mg / kg) 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.

EXAMPLE OF 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 average arterial pressure (SRAD) 53.9 ± 2.7 mm Hg, while 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 using daily, for 7 days intraperitoneal injection of L-NAME (N-nitro-L-arginine methyl ester at a dose of 25 mg / kg) caused arterial hypertension (SBP - 190.3 ± 6.7, DBP - 145.0 ± 3.9, СРАД - 160.1 ± 4.6 mm Hg) and led to a smaller decrease in blood pressure after administration of AH (SBP to 110.6 ± 5.2, DBP to 82.8 ± 6.6, the control system up to 92.1 ± 6.1 mm Hg) and the 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 enalapril (0.5 mg / kg intragastrically) did not lead to an optimal decrease in the initial BP values (SBP - 183.9 ± 11.4, DBP - 138.9 ± 7.0 mm Hg). ), and in experiments where resveratrol was still additionally administered (2 mg / kg intraperitoneally), potentiation of the hypotensive effect was noted, since the initial numbers of blood pressure were significantly lower: SBP - 175.9 ± 13.2, DBP - 132.6 ± 10, 6 mmHg (Table 1).

Table 1 Dynamics of blood pressure and heart rate in modeling nitric oxide deficiency and correction of endothelial dysfunction with enalapril using resveratrol Groups of animals Functional Tests GARDEN, mmHg DBP, mmHg Art. Heart rate, beats in min Intact Source 137.7 ± 3.7 101.9 ± 4.3 420.0 ± 9.0 EZV with AH 84.3 ± 4.5 38.7 ± 2.8 416.0 ± 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 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) + Enalapril (0.5 mg / kg) Source 183.9 ± 11.4 * 138.9 ± 7.0 * 377.0 ± 9.0 EZV with AH 98.0 ± 3.3 * 61.6 ± 4.7 * 381.0 ± 11.0 ENZV with NP 88.7 ± 4.0 42.7 ± 3.3 381.0 ± 11.0 L-NAME + Enalapril (0.5 mg / kg) + Resveratrol (2 mg / kg) Source 175.9 ± 13.2 ** 132.6 ± 10.6 ** 374.0 ± 12.0 EZV with AH 85.0 ± 7.5 ** 53.0 ± 5.7 ** 326.0 ± 14.0 ENZV with NP 84.8 ± 7.8 46.7 ± 3.0 378.0 ± 11.0 * - p <0.05 in comparison with the group of intact ** - p <0.05 compared with the L-NAME + enalapril group

Potentiation of a decrease in blood pressure in response to the administration of AH was also noted. The fundamental difference in ESV and ENZV in intact animals and animals with the introduction of an inhibitor of NO synthase L-NAME naturally led us to the necessity of deriving a special indicator characterizing the degree of endothelial dysfunction, which is the ratio of the area of the triangle over the trend in the reaction of restoration of blood pressure in response to the introduction of NP to the area triangle above the trend of the reaction of blood pressure recovery in response to the introduction of AH.

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

Thus, for a further assessment of the effect of AH and NP in ESV and ENZV, we used this ratio as an indicator of the correction of endothelial dysfunction. So, in the group where enalapril was administered only against the background of the administration of the L-NAME synthase inhibitor L-NAME, this ratio corresponded to a value of 3.2 ± 0.1, and in the group with additional administration of resveratrol to enalapril, 1.5 ± 0.1 ( table 2).

table 2 Indicators reflecting the correction of endothelial dysfunction with the administration of L-NAME enalapril and enalapril in combination with resveratrol 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 AH to NP L-NAME + Enalapril OH 80.1 ± 5.5 35.0 ± 1.7 1405.5 ± 97.5 3.2 ± 0.1 NP 109.0 ± 8.0 98.7 ± 5.6 5123.9 ± 121.8 L-NAME + Enalapril + Resveratrol OH 76.0 ± 8.6 46.3 ± 4.5 1742.2 ± 165.2 1.5 ± 0.1 * NP 79.0 ± 6.0 71.5 ± 7.6 2329.1 ± 185.3 * - p <0.05 compared with the L-NAME + enalapril group

Thus, the results obtained indicate the activation of the correction of endothelial dysfunction of enalapril in combination with its use with resveratrol.

Claims (1)

A method for correcting endothelial dysfunction, including the administration of enalapril, characterized in that the experiment simulates dysfunction by daily, for 7 days, intraperitoneal administration of L-nitro-arginine methyl ether to Wistar rats at a dose of 25 mg / kg, assessing the degree of development of dysfunction by the ratio of endothelium-independent and endothelium-dependent vasodilation, and against the background of this, enalapril is administered intragastrically at a dose of 0.5 mg / kg and resveratrol intraperitoneally at a dose of 2 mg / kg once a day.