RU2438188C1 - Method of endothelial dysfunction correction by combination of atorvastatin and resveratrol in l-name induced nitrogen oxide deficiency - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: endothelial dysfunction is ensured by intragastric introduction to a laboratory animal of the NO-synthase inhibitor - N-nitro-L-arginine of methyl ester (L-NAME) 25 mg/kg daily for 7 days. The endothelial dysfunction correction is ensured by intragastric introduction of atorvastatin 2.2 mg/kg and intragastric introduction of resveratrol 2 mg/kg once a day.

EFFECT: effective action of the combination of two specified preparations on vascular endothelium function.

1 ex, 2 tbl

Description

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

Closest to the claimed solution is a method for correcting endothelial dysfunction with atorvastatin in the presence of L-NAME, described by Lorkowska B, Chlopicki S. in the article "Statins as coronary vasodilators in isolated bovine coronary arteries-participation of PGI2 and NO" Prostaglandins Leukot Essent Fatty Acids, 2005 Feb 72 (2): 133-8.

The main disadvantage of this method is that the correction of endothelial dysfunction is associated with the pleiotropic effects of statins by activation of the gene for the expression of endothelial NO synthase (eNOS) and anti-inflammatory effects. Therefore, the results of the correction of endothelial dysfunction due to damage in the L-arginine-eNOS-NO system, which is mainly associated with a decrease in the activity and "uncoupling" of endothelial NO synthase due to oxidative stress with only atorvastatin, are unsatisfactory.

The objective of the invention is a method for the correction of endothelial dysfunction, including the use of atorvastatin, an inhibitor of HMG-CoA reductase and an activator of NO synthase, an antioxidant resveratrol.

This object is achieved by the fact that, against the background of modeling endothelial dysfunction in the experiment, an intraperitoneal injection of 25 mg / kg NO blocker for the synthesis of NO L-nitro-arginine methyl ether (L-NAME) into the laboratory animal is corrected by intragastric administration of atorvastatin in a dose of 2.2 mg / kg once a day and intraperitoneal administration of an activator of NO synthase of resveratrol at a dose of 2 mg / kg also once a day, which leads to activation of the correction of endothelial dysfunction, since resveratrol is tsya activator NO-synthase enzyme and antioxidant, by which NO produced in vascular endothelium and increased bioavailability.

The method is carried out as follows.

The experiments were carried out on white Wistar male rats 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 8th day from the start of the experiment, under anesthesia (chloral hedrate 300 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 TSD104A sensor and the MP 100 hardware and software complex, manufactured by Viopac System, Inc., USA. 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 PERFORMANCE

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) of 84.3 ± 4.4, for diastolic pressure (DBP) of 38.7 ± 2.8 and for average arterial pressure (SAD) 53.9 ± 2.7 mm Hg, while in 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 intraperitoneal injection of L-NAME (N-nitro-L-arginine methyl ether at a dose of 25 mg / kg) caused arterial hypertension (SBP 190.3 ± 6.7, DBP 145 , 0 ± 3.9, СРД 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, SRAD up to 92.1 ± 6.1 mm Hg) and NP (SBP up to 88.7 ± 4.7, DBP up to 50.8 ± 4.2 and SRAD up to 63.4 ± 4.1 mm Hg) compared with intact animals.

The simultaneous administration of L-NAME and atorvastatin (2.2 mg / kg intragastrically) led to a decrease in the initial values of blood pressure (SBP 162.8 ± 20.9, DBP 117.5 ± 14.4 mm Hg), and In experiments where resveratrol was additionally administered (2 mg / kg ip), a hypotensive effect was also observed, since the initial blood pressure values were lower: SBP 177.3 ± 14.0, DBP 137 ± 9.6 mm Hg. (table 1).

Table 1 Dynamics of blood pressure and heart rate in modeling nitric oxide deficiency and correction of endothelial dysfunction with atorvastatin using resveratrol Groups of animals Functional Tests GARDEN, mmHg DBP, mmHg 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 ± 11.0 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) + Atorvastatin (2.2 mg / kg) Source 162.8 ± 20.9 117.5 ± 14.4 317.5 ± 26.2 EZV with AH 96.3 ± 9.9 57.2 ± 4.9 303.3 ± 28.4 ENZV with NP 96.5 ± 13.7 53.0 ± 6.4 352.2 ± 36.7 L-NAME + Atorvastatin (2.2 mg / kg) + Resveratrol (2 mg / kg) Source 177.3 ± 14.0 ** 137.7 ± 9.6 ** 356.0 ± 20.5 EZV with AH 82.3 ± 8.6 ** 56.7 ± 6.2 ** 324.0 ± 18.6 ENZV with NP 89.1 ± 7.0 63.3 ± 5.8 358.4 ± 21.1 * - p <0.05 in comparison with the group of intact ** - p <0.05 compared with the L-NAME + atorvastatin group

Potentiation of a decrease in blood pressure in response to the management 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 restoration of blood pressure 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 only atorvastatin was administered against the background of administration of the NO synthase inhibitor L-NAME, this ratio corresponded to a value of 2.8 ± 0.5, and in the group with additional administration of resveratrol to atorvastatin, 2.1 ± 0.4 ( table 2).

Thus, the results obtained indicate the activation of the correction of endothelial dysfunction with atorvastatin with its combined use with resveratrol.

Claims (1)

A method for correcting endothelial dysfunction, including the use of atorvastatin, characterized in that in an experiment for a laboratory animal with endothelial dysfunction, against the background of its simulation by intraperitoneal administration of N-nitro-L-arginine methyl ester (L-NAME) at a dose of 25 mg / kg for 7 days, atorvastatin is administered intragastrically at a dose of 2.2 mg / kg and resveratrol is administered intraperitoneally at a dose of 2 mg / kg once a day.