RU2367421C1 - Method of lamotrigine correction of endothelial dysfunction associated with 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 that is ensured by simulation of endothelial dysfunction in experiment by daily intraperitoneal introduction to Wistar male rats N-nitro-L-apramiH methyl ester in a dose 25 mg/kg within 7 days. Correction of the dysfunction is enabled by intragastric introduction of Lamotrigine in a dose 8.5 mg/kg once a day.

EFFECT: method ensures possibility for correcting the endothelial dysfunction with antiepileptic agent Lamotrigine previously unknown as such.

2 tbl

Description

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

Closest to the claimed solution is a method for correcting systemic endothelial dysfunction by using a selective inhibitor of neuronal NO synthase 7-nitro-inidazole, which has anticonvulsant properties under conditions of oxidative stress in rats, described by N.E. Maksimovich in the work “The role of various isoforms of no synthases in endothelial damage and the formation of its dysfunction in brain ischemia in rats / N.E. Maksimovich // Belarusian Medical Journal. - 2004. - No. 3. - P.65 - 68, which consists in the intravenous administration of 7-N1 at a dose of 10 mg / kg to anesthetized animals when modeling oxidative stress by occlusion of the common carotid arteries. The vasoactive effects of the endothelium of the systemic vessels of rats were studied by examining the EDVD of the aortic rings in response to acetylcholine and the ENVD of the aortic rings in response to glycerol trinitrate.

The main disadvantage of this method is the lack of assessment of the functional state of the vascular bed in a holistic organism when modeling a deficiency of nitric oxide in the experiment and the lack of data on the widespread clinical use of 7-NI compared to lamotrigine.

The technical result of the invention is a method for the correction of endothelial dysfunction with L-NAME-induced nitric oxide deficiency, including the use of lamotrigine. The technical result is achieved by the fact that, against the background of modeling endothelial dysfunction in the experiment, an intraperitoneal daily administration to the laboratory animal of the synthesis inhibitor of NO N-nitro-L-arginine methyl ester (L-NAME) at a dose of 25 mg / kg for 7 days is carried out by daily correction intragastric administration of lamotrigine at a dose of 8.5 mg / kg

The method is carried out as follows.

The experiments were carried out on white rats male Wistar line weighing 250-300 g. N-nitpo-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 Biopac 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 s 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 an average arterial pressure (SAD) of 53.9 ± 2.7 mm Hg, while during the first 2-3 s a 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 sec. Blockade of NO synthase by means of prolonged, daily, intraperitoneal administration 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 (SBP to 110.6 ± 5.2, DBP to 82 , 8 ± 6.6, SAD up to 92.1 ± 6.1 mm Hg) and NP (SAD up to 88.7 ± 4.7, DBP up to 50.8 ± 4.2 and SAS up to 63.4 ± 4.1 mmHg) compared with intact animals.

The simultaneous administration of L-NAME and lamotrigine did not lead to a decrease in the initial values of blood pressure (table 1).

Table 1. Dynamics of blood pressure and heart rate in modeling nitric oxide deficiency and correction of endothelial dysfunction with lamotrigine. Groups of animals Functional Tests GARDEN, mmHg Art. 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 EZVsAH 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) + lamotrigine (8.5 mg / kg) Source 201.2 ± 12.7 146.4 ± 8.6 360.3 ± 13.9 EZVsAH 95.3 ± 4.0 54.4 ± 3.9 ** 332.1 ± 26.1 ENZV with NP 101.7 ± 3.7 ** 51.6 ± 3.9 356.9 ± 12.9 * - p <0.05 in comparison with the intact group; ** - p <0.05 compared with the L-NAME group

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, to further evaluate the effect of AH and NP in ESV and ENZV, we used this ratio as an indicator of correction of endothelial dysfunction. So, in the group, where lamotrigine was introduced against the background of the administration of the NO synthase inhibitor L-NAME, this ratio corresponded to a value of 1.9 ± 0.2 (table 2).

Table 2. Indicators reflecting the correction of endothelial dysfunction with the introduction of L-NAME lamotrigine. 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 Intact OH 59.9 ± 2.9 42.2 ± 0.8 1268.0 ± 74.8 1.1 ± 0.1 NP 61.0 ± 3.0 45.1 ± 1.0 1375.3 ± 93.7 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 + lamotrigine 8.5 mg / kg OH 74.3 ± 13.1 37.4 ± 3.3 ** 1389.2 ± 274.2 ** 1.9 ± 0.2 ** * - p <0.05 compared to the intact group, ** - p <0.05 compared to the L-NAME group

Thus, the results obtained allow us to ascertain the correction of endothelial dysfunction with lamotrigine during administration of L-NAME.

Claims (1)

A method for correcting endothelial dysfunction, characterized in that in an experiment in white rats, males of the Wistar strain with endothelial dysfunction amid its modeling by intraperitoneal administration of an inhibitor of endothelial synthase N-nitro-L-arginine methyl ester at a dose of 25 mg / kg daily for 7 days assessing the degree of development of dysfunction in relation to the indices of endothelium-independent and endothelium-dependent vasodilation, lamotrigine 8.5 mg / kg is administered intragastrically daily for 7 days.