RU2438189C1 - Method for hypoestrogen-induced endothelial dysfunction correction with enalapril in complex 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 daily intragastric introduction of enalapril 0.5 mg/kg for 6 weeks following the ovariectomy. The mixed solutions of homoeopathic dilutions of polyclonal rabbit C12, C30, C200 antibodies to human endothelial nitrogen oxide synthase are also introduced. 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 for the correction of hypoestrogen-induced endothelial dysfunction with furostanol glycosides from the cell culture of the plant dioscorea deltoidea, described on 27.07. 2008 in the patent of the Russian Federation 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 nitric oxide synthase - 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 main mechanism of action of enalapril in endothelial dysfunction is associated with activation of the expression of endothelial NO synthase, which also leads to the correction of endothelial dysfunction.

The objective of the invention is a method for the correction of hypoestrogen-induced endothelial dysfunction, including the use of enalapril, an ACE inhibitor, and a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200.

This object is achieved in that, against the background of modeling endothelial dysfunction in an experiment using bilateral ovariectomy in laboratory animals, it is corrected for 6 weeks by intragastric administration of enalapril at a dose of 0.5 mg / kg once a day and daily administration 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 quantities accepted in homeopathy, with free access to drinking, water replacement in drinking bowls it was made 1 time per day with quantitative accounting of the drunk liquid (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 and 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) - nutrient administration of sodium nitroprusside (NP) at a dose of 30 μg / kg.

The degree of development of endothelial dysfunction is estimated using the coefficient of endothelial dysfunction (QED), which is the ratio of the area of the triangle above the trend of the reaction of blood pressure recovery in response to the introduction of NP to the area of the triangle above the trend of the reaction of blood pressure recovery in response to the introduction of AH.

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 83.9 ± 4.8 mm Hg, for diastolic pressure (DBP) - 41, 9 ± 2.2 mm Hg and for average arterial pressure (Cf. HELL) 55.8 ± 2.2 mm Hg, with a sharp bradycardia developing up to 130-150 beats per minute during the first 2-3 seconds (Table 1).

Recovery of blood pressure occurred on average for 43.6 ± 2.2 s 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 Cf. Blood pressure up to 62.7 ± 3.2 mm Hg (Table 1), followed by complete recovery on average within 40.2 ± 2.4 s (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.

Processing of the obtained experimental data during functional tests for endothelium-dependent (acetylcholine 40 μg / kg iv) and endothelium-independent (nitroprusside 30 mg / kg iv) relaxation of vessels in experimental animals allowed us to establish that QED in rats of the intact group was 0.8 and rats after modeling hypoestrogen-induced nitric oxide deficiency are 2.5 times more - 2.1.

Table 1 Dynamics of blood pressure and heart rate in modeling hypoestrogen-induced nitric oxide deficiency and its correction with enalapril in combination with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial nitric oxide synthase - C12, C30, C200. Groups of animals Functional Tests GARDEN, mmHg DBP, mmHg Heart rate, beats in minutes 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 condition + enalapril Source 139.6 ± 7.3 ** 107.4 ± 4.9 ** 397.4 ± 12.2 OH 80.7 ± 4.6 41.9 ± 2.7 ** 378.7 ± 9.9 NP 81.6 ± 5.8 40.1 ± 4.7 ** 379.5 ± 13.4 Hypoestrogenic state + enalapril + mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to human endothelial NOS - C12, C30, C200 Source 150.6 ± 9.6 * 111.4 ± 6.7 ** 385.4 ± 14.0 EZV with AH 104.4 ± 12.9 * 62.4 ± 3.9 * 378.4 ± 14.8 ENZV with NP 113.6 ± 8.8 ** 72.4 ± 7.7 ** 385.6 ± 11.0 * - p <0.05 in comparison with the intact group; ** - p <0.05 compared with the group with hypoestrogenic state.

Intragastric administration of enalapril at a dose of 0.5 mg / kg once a day and adding to the drinkers with free access to drink a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide - C12, C30, C200 (BD Transduction Labs, USA) led to a pronounced decrease in the indicators of the initial blood pressure values (SBP - 150.6 ± 9.6, DBP - 111.4 ± 6.7 mm Hg) (Table 1.).

However, in the group where enalapril and a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to human endothelial synthase of human nitric oxide - C12, C30, C200, QED were administered for 6 weeks against the background of bilateral oophorectomy, it was 0.9, which is significantly lower than in the group with hypoestrogenic state. Whereas in the group with enalapril monotherapy, this indicator was 1.3 (Table 2.)

table 2 Indicators reflecting the correction of hypoestrogen-induced endothelial dysfunction on the background of bilateral ovariectomy with enalapril in combination with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to human nitric oxide endothelial synthase - 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, conventional 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 condition + enalapril OH 63.3 ± 4.2 49.0 ± 2.2 ** 1573.6 ± 82.0 ** 1.3 ± 0.1 ** NP 75.5 ± 9.5 59.4 ± 3.5 ** 1984.0 ± 262.2 * Hypoestrogenic state + enalapril + mixture of solutions of homeopathic dilutions of polyclonal antibodies to human endothelial NOS - C12, C30, C200 OH 48.1 ± 5.7 * 60.4 ± 3.0 ** 1468.1 ± 214.8 ** 0.9 ± 0.1 ** NP 43.6 ± 4.4 ** 60.4 ± 2.2 ** 1327.8 ± 168.1 ** * - 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 enalapril in combination with a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to human endothelial synthase nitric oxide - C12, C30, C200, reducing the CED to the level of animals in animals.

Claims (1)

A method for the correction of hypoestrogen-induced endothelial dysfunction, characterized in that in the experiment, female Wistar rats are injected with enalapril at a dose of 0.5 mg / kg daily intragastrically for 6 weeks after ovariectomy and a solution mixture is added once a day to drinkers for drinking homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase of human nitric oxide - C12, C30, C200.