RU2343927C1 - Method of correction of endothelial dysfunction using lozartan in complex with admixture of solutions of homeopathic delutions of polyclonal rabbit antibodies to endothelial synthase of human nitrogen oxide-c12, c30, c200 - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: for correction of endothelial dysfunction in experiment to white rats to males of line Wistar with endothelial dysfunction, against its modelling by intraperitoneal introduction of an inhibitor endothelial synthase "M-nitro-L-arginine of methyl ether in a dose of 25 mg/kg within 7-days enter intragastrically lozartan in a dose of 6 mg/kg daily and an admixture of homeopathic solutions in delution of polyclonal rabbit antibodies to endothelial synthase of human nitrogen oxide - C12, C30, C200 addition into drinking bowls to drink.

EFFECT: effective correction of endothelial dysfunction.

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 using losartan, which is an angiotensin II receptor antagonist (AT 1 subtype), described by De Gennaro Colonna V. et al. "Angiotensin II type 1 receptor antagonism improves endothelial vasodilator function in L-NAME-induced hypertensive rats by a kinin-dependent mechanism", J. Hypertens. 2006 Jan; 24 (1): 95-102, which consists in the fact that male Sprague-Dawley rats were simulated endothelial dysfunction by introducing the N (ω) -nitro-L-arginine methyl ester (L-NAME) NO synthase blocker into drinking water for 8 weeks and studied the endothelioprotective effects of losartan (10 mg / kg / day), administered in the last 2 weeks of taking L-NAME, realized through the activation of bradykinin B2 receptors.

The main disadvantage of this method is the indirect mechanism of action of losartan in endothelial dysfunction, associated with stimulation of the AT 2 subtype of angiotensin II receptors and realized as a result of the bradykinin-dependent pathway of endothelioprotection, while damage to system L-arginine NO, leading to disruption of the production of nitric oxide (NO), the activity of which when using only losartan tsya not sufficiently compensated. While the main mechanism of action of a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200 is an increase in the number of eNOS, leading to an increase in the content of nitric oxide in the main vascular bed and correction of endothelial dysfunction.

Therefore, the results of the correction of endothelial dysfunction with a lack of NO synthases using losartan are unsatisfactory.

The technical result of the invention is a method for the correction of endothelial dysfunction, including the use of losartan, which is an antagonist of angiotensin II receptors (AT 1-subtype) and a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide - C12, C30, C200.

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 for the synthesis of NO L-nitro-arginine methyl ether (L-NAME) at a dose of 25 mg / kg is carried out by intraperitoneal administration of losartan in a laboratory animal for 7 days a dose of 6 mg / kg once a day and the introduction of a mixture of homeopathic dilutions of affinity-purified antibodies to endothelial NOS - C12, C30, C200 at the rate of 1 tablet per 100 ml of water with free access to drink, the water in the drinkers was replaced 1 time per day with quantitative consideration of the drunk liquid (20 ± 3 ml on average per animal), which leads to an increase in the content of nitric oxide in the main vascular bed and correction of endothelial dysfunction.

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, under anesthesia (ethaminamin 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.

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) 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, 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 (SBP to 110.6 ± 5.2, DBP to 82, 8 ± 6.6, CAP up to 92.1 ± 6.1 mm Hg) and NP (CAP up to 88.7 ± 4.7, DBP up to 50.8 ± 4.2 and CAP up to 63.4 ± 4.1 mmHg) compared with intact animals.

The simultaneous administration of L-NAME and losartan (6 mg / kg intragastrically) did not lead to a decrease in the initial BP values (SBP - 192.2 ± 10.5, DBP - 138.2 ± 2.4 mm Hg). In experiments where a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide — C12, C30, C200 — was added to drinkers with free access to drinking, this did not lead to a significant decrease in the initial BP values (GARDEN - 185.4 ± 13.5, DBP - 138.1 ± 7 ± 9 mm Hg) (table 1).

Table 1
Dynamics of blood pressure and heart rate during modeling nitric oxide deficiency and correction of endothelial dysfunction with losartan when combined with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200 (n = 10, M ± m) . Groups of animals Functional Samples 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 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) + lozartan (6 mg / kg) Source 192.2 ± 10.5 * 138.2 ± 2.4 * 381.0 ± 9.0 EZV with AH 90.0 ± 7.8 * 51.7 ± 5.0 * 365.0 ± 8.0 ENZV with NP 95.5 ± 8.9 55.5 ± 5.5 378.0 ± 12.0 L-NAME (25 mg / kg) + lo-zartan +
a mixture of solutions of homeopathic dilutions
of rabbit antibodies to human endothelial NOS - C12, C30, C200 Source 185.4 ± 13.5 138.1 ± 7.9 390.8 ± 5.1 EZV with AH 65.3 ± 10.2 ** 37.6 ± 5.9 ** 371.1 ± 5.2 ENZV with NP 97.3 ± 6.3 55.7 ± 6.1 402.1 ± 4.1 * p <0.05 in comparison with the intact group; ** - p <0.05 compared with the L-NAME + losartan group
Note: CAD - systolic blood pressure, DBP - diastolic blood pressure, heart rate - heart rate, ESV - endothelium-dependent vasodilation, ENZV - endothelium-independent vasodilation, AH - acetylcholine, NP - nitroprusside

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 losartan was administered against the background of the administration of the NO synthase inhibitor L-NAME, this ratio corresponded to a value of 2.3 ± 0.3, and in the group with the addition of lasartan with a mixture of solutions of homeopathic dilution of polyclonal rabbit antibodies to endothelial synthase human nitric oxide - C12, C30, C200 - 1.5 ± 0.2 (table 2)

table 2
Indicators reflecting the correction of endothelial dysfunction during administration of L-NAME with losartan when combined with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200 (n = 10, M ± m). Groups of animals Functional Samples 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 AHKNP L-NAME + Lo Zartan OH 91.7 ± 4.7 29.7 ± 1.9 1048.9 ± 79.8 2.3 ± 0.3 NP 93.0 ± 7.0 74.8 ± 8.6 3523.4 ± 535.9 L-NAME + Lo Zartan +
a mixture of solutions of homeopathic dilutions of polyclonal antibodies to human endothelial NOS - C12, C30, C200 OH 107.1 ± 9.8 * 34.9 ± 1.2 * 1891.2 ± 202.4 * 1.5 ± 0.2 * NP 86.0 ± 7.1 * 59.2 ± 2.0 * 2587.8 ± 255.8 * * - p <0.05 compared with the group L-NAME + losartan
Note: AH - acetylcholine, NP - nitroprusside.

Thus, the obtained results allow us to state the correction of endothelial against the background of L-NAME administration with losartan when combined with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200.

Claims (1)

A method for the correction of endothelial dysfunction, including the use of losartan, an angiotensin II receptor antagonist (AT 1-subtype), characterized in that in the experiment to white rats Wistar males with endothelial dysfunction, against the background of its modeling by the intraperitoneal administration of an M-nitro endothelial synthase inhibitor -L-arginine methyl ester at a dose of 25 mg / kg for 7 days is administered intragastrically losartan at a dose of 6 mg / kg daily and a mixture of solutions of homeopathic dilution of polyclonal rabbit antibodies to endothelium ialnoy human nitric oxide synthase - C 12, C30, C200 adding to drinking troughs.