RU2421822C2 - Method of endothelial dysfunction correction by combination of trimetazidine and l-arginine in l-name-induced nitrogen oxide deficiency - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: endothelial dysfunction correction is ensured by simulating endothelial dysfunction by the intraperitoneal introduction to laboratory male rats Wistar of N-nitro-L-arginine methyl ether 25 mg/kg daily for 7 days. A development degree of endothelial dysfunction is estimated by the relation of endothelium-independent and endothelium-dependent vasodilation indicators. The endothelial dysfunction correction is ensured by the intragastric introduction of trimetazidine 6 mg/kg and the intraperitoneal introduction of L-arginine 200 mg/kg.

EFFECT: method provides activation of the L-NAME-induced endothelial dysfunction correction ensured by the introduction of a specific combination of the pharmacological preparations promoting the improvement of endothelial vasorelaxation properties.

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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 endothelial function using trimetazidine related to drugs that improve myocardial metabolism, described by Di Napoli P, Chierchia S, Taccardi AA, Grilli A, Felaco M, De Caterina R, Barsotti A in the article "Trimetazidine improves post-ischemic recovery by preserving endothelial nitric oxide synthase expression in isolated working rat hearts ”, Nitric oxide: biology and chemistry / official journal of the Nitric Oxide Society 2007 Mar; 16 (2): 228-36, which has a cardioprotective effect in L-NAME-induced nitric oxide deficiency in a model of experimental rat ischemia-reperfusion.

The main disadvantage of this method is the mediated mechanism of action of trimetazidine in endothelial dysfunction associated with the prevention of a decrease in the intracellular ATP content by maintaining the energy metabolism of cells in a state of hypoxia, ensuring the normal functioning of membrane ion channels, transmembrane transfer of potassium and sodium ions and preserving cellular homeostasis, while one of the main factors in the development of this pathophysiological condition, especially with essential ar terial hypertension, is damage in the L-arginine-NO system, leading to a violation of the basal release of nitric oxide (NO), which is not corrected with the use of only one trimetazidine. Therefore, the results of the correction of endothelial dysfunction with damage in the L-arginine-NO system, which is mainly associated with the deficiency of endothelial NO synthase, with the help of trimetazidine are unsatisfactory.

The technical result of the invention is a method for the correction of endothelial dysfunction, including the use of trimetazidine, related to drugs that improve myocardial metabolism, and a donor of NO - L-arginine.

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 the synthesis of NO N-nitro-L-arginine methyl ester (L-NAME) at a dose of 25 mg / kg for 7 days is performed to correct endothelial dysfunction for account of intragastric administration of trimetazidine at a dose of 6 mg / kg once a day and intraperitoneal administration of a donor NO L-arginine at a dose of 200 mg / kg once a day, which leads to activation of the correction of endothelial dysfunction, since L-argi Nin is the main substrate for the synthesis of NO and at the same time an activator of its enzymatic pathway of formation 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 a sensor and the Biopac Systems, Inc. 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 PERFORMANCE

A bolus intravenous administration of AH for 3-5 s 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) - 38.7 ± 2.8 and for average arterial pressure (SRAD) 53.9 ± 2.7 mm Hg, while during the first 2-3 s sharp bradycardia developed up to 130-150 beats per minute. Recovery of blood pressure occurred on average for 42.2 ± 0.8 s 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 s. Blockade of NO synthase with the help of a long, 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 - 169.4 ± 6.3, DBP - 131.4 ± 5.0, SAD - 144.1 ± 4.4 mm Hg) and led to a lower decrease in blood pressure after administration of AH (SBP to 100.6 ± 8.1, DBP to 68, 8 ± 8.0, a control system up to 79.4 ± 6.1 mmHg) and an NP (a control system up to 90.7 ± 4.7, a DBP up to 61.4 ± 4.2 and a control system up to 71.2 ± 4.1 mmHg) compared with intact animals.

The simultaneous administration of L-NAME and trimetazidine (6 mg / kg intragastrically) did not lead to a decrease in the initial BP values (SBP - 162.6 ± 4.5, DBP - 132.3 ± 4.1 mm Hg), and in experiments where L-arginine was additionally introduced (200 mg / kg intraperitoneally), potentiation of the hypotensive effect was noted: GARDEN - 152.7 ± 1.4 DBP -123.3 ± 1.4 mm Hg. (Table 1).

Table 1 Dynamics of blood pressure and heart rate in modeling nitric oxide deficiency and correction of endothelial dysfunction with a combination of trimetazidine and L-arginine Groups of animals Functional Tests GARDEN, mmHg DBP, mmHg Heart rate, beats in minutes 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 169.4 ± 6.3 * 131.4 ± 5.0 * 432.0 ± 9.0 EZV with AH 100.6 ± 8.1 68.8 ± 8.0 * 426.0 ± 13.0 ENZV with NP 90.7 ± 4.7 61.4 ± 4.2 * 423.0 ± 14.0 L-NAME (25 mg / kg) + trimetazidine (6 mg / kg) Source 162.6 ± 4.5 * 132.3 ± 4.1 * 346.5 ± 14.3 EZV with AH 99.8 ± 3.6 * 58.1 ± 2.0 * 319.6 ± 16.8 ENZV with NP 95.4 ± 6.6 50.5 ± 2.4 367.9 ± 12.5 L-NAME (25 mg / kg) + trimetazidine (6 mg / kg) + L-arginine (200 mg / kg) Source 152.7 ± 1.4 * ^y 123.3 ± 1.4 * ^y 375.6 ± 19.1 EZV with AH 94.0 ± 5.1 57.6 ± 5.9 * 361.7 ± 18.9 ENZV with NP 89.1 ± 9.3 57.0 ± 4.7 * 376.9 ± 15.0 ∗ - p <0.05 compared with the group of intact ^y - p <0.05 compared with the group L-NAME + trimetazidine

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 need to derive 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 (endothelium-independent vasodilation) to the area of the triangle above the trend of the blood pressure restoration reaction in response to the administration of AH (endothelium-dependent vasodilation).

We calculated this ratio in each animal of the intact group and rats after modeling the blockade of NO synthase and obtained a difference of this indicator by 4.5 times - 1.1 ± 0.1 in intact and 4.9 ± 0.6 in animals treated, respectively L-NAME.

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 trimetazidine was administered against the background of administration of the L-NAME synthase inhibitor L-NAME, this ratio corresponded to a value of 4.2 ± 0.6, and in the group with additional administration of the NO donor of L-arginine to trimetazidine, 2.5 ± 0.2 (table 2).

table 2 Indicators reflecting the correction of endothelial dysfunction during administration of L-NAME with trimetazidine and a combination of trimetazidine and L-arginine Groups of animals Functional Tests The increase in the fall of the vascular reaction by SRAD (mm Hg) Vascular reaction time (s) The area of the vascular reaction (conventional units) The ratio of the areas of the vascular reaction of AH to NP L-NAME (25 mg / kg) + trimetazidine (6 mg / kg) OH 70.4 ± 4.6 33.3 ± 5.3 1163.1 ± 176.7 4.2 ± 0.6 NP 90.0 ± 5.0 98.4 ± 8.2 4462.8 ± 444.9 L-NAME (25 mg / kg) + trimetazidine (6 mg / kg) + L-arginine (200 mg / kg) OH 72.3 ± 5.5 54.4 ± 4.8 ^y 2000.5 ± 276.6 ^y 2.5 ± 0.2 ^y NP 97.0 ± 8.0 99.4 ± 7.4 4870.0 ± 643.1 ^y - p <0.05 compared with the group L-NAME + trimetazidine

Thus, the results obtained allow us to ascertain the activation of correction of endothelial dysfunction against the background of the introduction of L-NAME by a combination of trimetazidine and L-arginine.

Claims (1)

A method for the correction of endothelial dysfunction, characterized in that in an experiment for laboratory rats male Wistar strain 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 the development of dysfunction in relation to the indices of endothelium-independent and endothelium-dependent vasodilation is administered a combination of trimetazidine intragastrically at a dose of 6 mg / kg and L-arginine intraperitoneally at a dose of 200 mg / kg.