RU2438191C1 - Method for hypoestrogen-induced endothelial dysfunction correction with mixed solutions of homoeopathic dilutions of polyclonal rabbit c12, c30, c200 antibodies to human endothelial nitrogen oxide synthase - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: for the purpose of hypoestrogen-induced endothelial dysfunction correction in white female Wistar rats, hypoestrogen-induced endothelial dysfunction is simulated by bilateral ovariectomy. The dysfunction correction is ensured by introduction of mixed solutions of homoeopathic dilutions of polyclonal rabbit C12, C30, C200 antibodies to human endothelial nitrogen oxide synthase for 6 weeks following the ovariectomy. The specified mixture is added to drinking bowls once a day.

EFFECT: method provides effective correction with reducing an endothelial dysfunction coefficient in females to the level observed in intact animals ensured by the effect on various developmental mechanisms of this dysfunction in hypooestrogenic conditions.

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Description

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

Closest to the claimed solution is a method of correction of hypoestrogen-induced endothelial dysfunction with furostanol glycosides from a cell culture of the plant dioscorea deltoidea, described on July 27, 2008 in RF patent No. 23239807.

The main disadvantage of this method is that the correction of hypoestrogen-induced endothelial dysfunction is associated with the estrogen-like activity of furostanol glycosides. Therefore, the results of the correction of endothelial dysfunction due to damage in the L-arginine-NO system, which is mainly associated with deficiency of endothelial NO synthase, using furostanol glycosides from a cell culture of the dioscorea deltoidea plant with estrogen-like activity are unsatisfactory.

While the main mechanism of action of a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200 is an increase in the number of eNOS, thereby affecting the main pathogenesis of the development of endothelial dysfunction, leading to an increase in the content of nitric oxide in the main vascular channel and correction of endothelial dysfunction.

The objective of the invention is a method of correction of hypoestrogen-induced endothelial dysfunction, including the use of a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200, leading to an increase in the number of eNOS.

This object is achieved by the fact that, against the background of modeling endothelial dysfunction in an experiment using bilateral ovariectomy in laboratory animals, it is corrected for 6 weeks by daily injection of a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of human nitric oxide - C12, C30, C200 by adding to drinking bowls in the quantities accepted in homeopathy, with free access to drink, the water in the drinking bowls was changed 1 time per day with quantitative consideration of the drunk liquid spine (20 ± 3 ml on average per animal).

The method is carried out as follows.

The experiments were performed on white Wistar female rats weighing 200-300 g. To simulate hypoestrogen-induced endothelial dysfunction, rats were anesthetized with sodium ethamine (50 mg / kg) and bilateral ovariectomy was performed. On day 42 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 TSD104A sensor and the MP100 hardware-software complex, manufactured by Biopac 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 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) of 83.9 ± 4.8 mm Hg, for diastolic pressure (DBP) - 41.9 ± 2.2 mm Hg and for average arterial pressure (Avg BP) 55.8 ± 2.2 mm Hg, while in the first 2-3 seconds a sharp bradycardia developed up to 130-150 beats per minute (Table 1).

Recovery of blood pressure occurred on average for 43.6 ± 2.2 seconds after normalization of the heart rhythm. It was this integral that we accepted as the vascular component of the reaction to AX and was used for further calculations (Table 2). ENZV was also characterized by a decrease in SBP to 91.0 ± 4.6, DBP to 48.5 ± 2.9 and Avg. BP to 62.7 ± 3.2 mm Hg. (Table 1) with subsequent complete recovery on average within 40.2 ± 2.4 seconds (Table 2).

The hypoestrogenic state, 6 weeks after ovariectomy, caused arterial hypertension (SBP - 160.0 ± 6.2, DBP - 124.9 ± 5.5, SRAD - 136.6 ± 5.6 mm Hg) and led to a smaller decrease in blood pressure after the administration of AH (SBP to 91.8 ± 8.0, DBP to 57.3 ± 7.1, SBP to 68.8 ± 7.3 mm Hg) and NP (SBP to 87.4 ± 7.5, DBP up to 57.8 ± 6.6 and CAPP up to 67.7 ± 6.1 mm Hg) compared with intact animals (Table 1).

After 6 weeks, after female Wistar rats undergoing bilateral ovariectomy, there was a significant decrease in the integral (reaction of blood pressure restoration after administration of AX) to 1022.5 ± 131 srvc. compared with the intact group of animals in which this indicator was 1673.6 ± 132.4 srvc. (table 1). On the contrary, the ESD, which is a reflection of the reaction to the administration of NP, increased in animals with experimental endothelial dysfunction and its integral was 2157.3 ± 167 conventional units, respectively. and 1338.1 ± 121.1 conventional units in the control group.

The fundamental difference in ESV and ENVS in intact animals and animals with a hypoestrogenic state naturally led us to the necessity of deriving a special indicator, the coefficient of endothelial dysfunction (QED), which characterizes the degree of endothelial dysfunction, which is the ratio of the area of the triangle over the trend of the reaction of blood pressure restoration in response to the introduction of NP to the area of the triangle above the trend of blood pressure recovery reaction in response to the administration of AH.

Table 1 Dynamics of blood pressure and heart rate during modeling of hypoestrogen-induced deficiency of nitric oxide and its correction with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200. Groups of animals Functional Tests GARDEN, mmHg DBP, mmHg Heart rate, beats in min Intact Source 128.1 ± 6.0 95.7 ± 4 401.0 ± 8.0 OH 83.9 ± 4.8 41.9 ± 2.2 398.0 ± 11.0 NP 91.0 ± 4.6 48.5 ± 2.9 402.0 ± 10.0 Hypoestrogenic condition Source 160.0 ± 6.2 * 124.9 ± 5.5 * 385.0 ± 13.0 OH 91.8 ± 8.0 57.3 ± 7.1 * 385.0 ± 9.0 NP 87.4 ± 7.5 * 57.8 ± 6.6 397.0 ± 8.0 Hypoestrogenic state + a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to human endothelial NOS - C12, C30, C200 Source 158.5 ± 15.0 120.7 ± 9.9 377.9 ± 18.0 EZV with AH 103.4 ± 10.9 60.3 ± 5.1 381.0 ± 16.4 ENZV with NP 119.6 ± 12.3 78.4 ± 8.2 379.9 ± 15.0 * - p <0.05 in comparison with the intact group; ** - p <0.05 compared with the group with hypoestrogenic state

We calculated this ratio for each animal in the intact group and rats after modeling a hypoestrogen-induced deficiency of nitric oxide and obtained a 2-fold difference in this indicator - 0.8 in intact and 2.1 in animals, respectively, with a hypoestrogenic state.

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.

The addition of a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of nitric oxide - C12, C30, C200 (BD Transduction Labs, USA) to drinkers with free access to drinking did not lead to a decrease in the initial BP values (GARDEN - 158.5 ± 15 , 0, DBP - 120.7 ± 9.9 mmHg) (Table 1.).

However, in the group where, against the background of bilateral ovariectomy, a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of human nitric oxide - C12, C30, C200 QED was administered for 6 weeks, it was 1.1, which was significantly lower than in the group with hypoestrogenic state .

table 2 Indicators reflecting the correction of hypoestrogen-induced endothelial dysfunction on the background of bilateral ovariectomy with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to human endothelial synthase nitric oxide - C12, C30, C200. 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) QED, conv.
units Intact OH 56.7 ± 4.1 40.5 ± 2.6 1673.6 ± 132.4 0.8 ±
0.1 NP 60.0 ± 4.8 58.3 ± 2.9 1338.1 ± 121.1 Hypoestrogenic condition OH 67.8 ± 7.3 * 30.0 ± 5.4 * 1022.5 ± 131 * 2.1 ±
0.2 * NP 79.2 ± 8.8 * 46.0 ± 4.5 * 2157.3 ± 167 * Hypoestrogenic state + a mixture of solutions of homeopathic dilutions of polyclonal antibodies to human endothelial NOS - C12, C30, C200 OH 65.1 ± 5.8 * 61.9 ± 4.3 ** 2015.0 ± 240.7 ** 1.1 ±
0,1 ** NP 50.5 ± 2.9 ** 82.0 ± 2.4 ** 2065.6 ± 116.9 * * - p <0.05 in comparison with the intact group, ** - p <0.05 in comparison with the group with hypoestrogenic state

Thus, the obtained results allow us to state the correction of hypoestrogen-induced endothelial on the background of bilateral ovariectomy with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of human nitric oxide - C12, C30, C200, reducing QED to the level of that in intact animals.

Claims (1)

A method for the correction of hypoestrogen-induced endothelial dysfunction, characterized in that in the experiment, female Wistar rats are injected daily once a day with 6 times after ovariectomy by adding a mixture of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of human nitric oxide - C12 to drinkers , C30, C200.