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

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.

2 tbl, 1 ex

 

The invention relates to medicine, in particular to the experimental cardiovascolari.

Closest to the claimed solution is a way of correcting hypoestrogen-induced endothelial dysfunction furostanol glycosides from cell culture plants of dioscorea deltoidea described 27.07. 2008 patent RF №2329807.

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

While the main mechanism of action of the mixture of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide - C12, C30, C200 is an increase of eNOS, thereby affecting the main link of pathogenesis of endothelial dysfunction, leading to increased levels of nitric oxide in the vascular trunk line and correction of endothelial dysfunction; the main mechanism of action of enalapril in endothelial dysfunction SV is associated with the activation of the expression of endothelial NO-synthase, which also leads to the correction of endothelial dysfunction.

Object of the invention is a method for correcting hypoestrogen-induced endothelial dysfunction, including the use of enalapril related to ACE inhibitors, and mixtures of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide - C12, C30, C200.

This object is achieved in that the background modeling of endothelial dysfunction in the experiment with bilateral oophorectomy in laboratory animals is its correction within 6 weeks intragastric introduction of enalapril in a dose of 0.5 mg/kg once per day and the daily introduction of the mixture of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide person - C12, C30, C200 adding to drinkers in quantities taken in homeopathy, with free access to drinking, replacement of water in the drinkers were performed 1 time per day with a quantitative accounting of fluid you drink (20±3 ml on average one animal).

The METHOD IS AS follows

The experiments were carried out on white rats female Wistar weighing 200-300 g For modeling hypoestrogen-induced endothelial dysfunction in rats was narcoticyou atenalol sodium (50 mg/kg) and held bilateral ovariectomy the Y. On day 42 from the beginning of the experiment under anesthesia (eleminal-sodium 50 mg/kg) catheter in the left carotid artery for recording blood pressure (BP), a bolus of pharmacological agents in the right femoral vein. Hemodynamic parameters: systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) was measured continuously by a sensor TSD104A and hardware-software complex MR production Biopac System, Inc., USA. Functional tests: endothelium-dependent vasodilatation (ESV) - intravenous administration of acetyl-choline (ACH) at a dose of 40 mcg/kg, endothelial-independent vasodilatation (ANSW) - nutrivene introduction of sodium nitroprusside (NP) at a dose of 30 µg/kg

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

When the statistical processing of the data was calculated the mean value, the standard deviation value. The differences were considered significant at p<0,05.

SPECIFIC EXAMPLE

Bolus intravenous AH 3-5 sec led to sharp the drop in blood pressure, reaching a peak in the intact animals for systolic blood pressure (SBP) of 83.9±4.8 mm Hg for diastolic blood pressure (DBP) - 41,9±2.2 mm Hg and mean arterial pressure (Cf. AD) 55,8±2.2 mm Hg, while during the first 2-3 sec developed sudden bradycardia up to 130-150 beats per minute (table 1).

Restoring the HELL happened in average 43,6±2.2 s after normalization of heart rate. This integral we have taken as the vascular component of the reaction on the AH and was used for further calculations (table 2). ANSW was also characterized by a decrease in the GARDEN to 91,0±4,6, dad to 48.5±2.9 and Cf. HELL to 62.7±3.2 mm Hg(table 1) followed by complete recovery within average 40,2±2,4 (table 2).

Hypoestrogenemia the state, within 6 weeks after oophorectomy, called arterial hypertension (GARDEN - 160,0±6,2, dad was 124.9±5,5, Srad - 136,6±5.6 mm Hg) and lead to a smaller decline in HELL after the introduction of AH (GARDEN up to 91.8±8,0, dad to 57.3±7,1, Srad to 68.8±7,3 mm Hg) and NP (the GARDEN to 87,4±7,5, dad to 57.8±6.6 and Srad to 67,7±6.1 mm Hg) compared with intact animals (table 1).

6 weeks after the females of Wistar rats with bilateral oophorectomy was a significant decrease of the integral (response and recovery actions after the introduction of the AH) to 1022,5±131 used compared with the intact group of animals in which the indicators of the ü was 1673,6±132,4 used (table 1). On the contrary, ENSR, reflecting the response to the introduction of the NP increased in animals with experimental endothelial dysfunction and its integral, respectively 2157,3±167 used and 1338,1±121, 1million used in the control group.

Processing of experimental data when carrying out functional tests on endothelium-dependent (acetylcholine 40 µg/kg) and endothelium-independent (nitroprusside 30 mg/kg/in) relaxation of blood vessels in experimental animals has allowed to establish that QED in rats intact group was 0.8, and rats after modeling hypoestrogen-induced deficiency of nitric oxide 2.5 times more - 2,1.

Table 1
Dynamics of blood pressure and heart rate when modeling hypoestrogen-induced deficiency of nitric oxide and its correction by enalapril in combination with the mixture of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide - C12, C30, C200.
Groups of animalsFunctional testsGARDEN, mm HgDBP, mm HgHeart rate, beats per min
Intact is e Source128,1±6,095,7±4401,0±8,0
OH83,9±4,841,9±2,2398,0±11,0
NP91,0±4,648,5±2,9402,0±10,0
Hypoestrogenic stateSource160,0±6,2*was 124.9±5,5*385,0±13,0
OHabout 91.8±8,057,3±7,1*385,0±9,0
NP87,4±7,557,8±6,6397,0±8,0
Hypoestrogenic state + enalaprilSource139,6±7,3**to 107.4±4,9**397,4±12,2
OH80,7±4,641,9±2,7**378,7±9,9
NP81,6±5,84,1±4,7** 379,5±13,4
Hypoestrogenic state + enalapril + blend solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial NOS man - C12, C30, C200Sourceto 150.6±9,6*byr111.4±6,7**385,4±14,0
ESV with AH104,4±12,9*62,4±3,9*378,4±14,8
ANSV with NP113,6±8,8**72,4±7,7**385,6±11,0
* - p<0,05 in comparison with the group intact; ** - p<0,05 in comparison with the group with hypoestrogenism state.

Intragastric administration of enalapril in a dose of 0.5 mg/kg once a day and add in drinkers with free access to drinking a mixture of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide - C12, C30, C200 (BD Transduction Labs, USA) did not lead to a pronounced decrease of the initial values of AD (GARDEN - to 150.6±9,6, DBP - byr111.4±6.7 mm Hg) (table 1).

However, in this group, where the background of bilateral oophorectomy for 6 weeks was administered enalapril and the mixture rastvorosmesiteli dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide person - C12, C30, C200, QED amounted to 0.9, which was significantly lower than in the group with hypoestrogenism condition. Whereas in the group with monotherapy with enalapril, this figure was 1.3 (table 2.)

Table 2
Indicators that reflect the correction hypoestrogen-induced endothelial dysfunction in the background of bilateral oophorectomy with enalapril in combination with the mixture of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide person - C12, C30 C200.
Groups of animalsFunctional testsThe increase in the incidence of vascular response to Srad (mm Hg)The vascular reaction time (sec.)The area of the vascular response (srvc. ed)QED, c.u
The intactOH56,7±4,140,5±2,61673,6±132,40,8±0,1
NP60,0±4,858,3±2,91338,1±121, 1million
Hypoestrogenic stateOH67,8±7,3*30,0±5,4*1022,5±131*2,1±0,2*
NP79,2±8,8*46,0±4,5*2157,3±167*
Hypoestrogenic state+enalaprilOH63,3±4,249,0±2,2**1573,6±82,0**1,3±0,1**
NP75,5±9,559,4±3,5**1984,0±262,2*
Hypoestrogenic state + enalapril + blend solutions of homeopathic dilutions of polyclonal antibodies to endothelial NOS man - C12, C30, C200OH48,1±5,7*60,4±3,0**1468,1±214,8**0,9±0,1**
NP43,6±4,4**60,4±2,2**1327,8±168,1**
* - p<0,05 in comparison with the group intact, ** - p<0,05 in comparison to the situation with the group with hypoestrogenism state.

Thus, the obtained results allow to conclude the correction hypoestrogen-induced endothelial amid bilateral oophorectomy with enalapril in combination with the mixture of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide person - C12, C30, C200, reducing QED to the level of that of intact animals.

The method for correcting hypoestrogen-induced endothelial dysfunction, characterized in that in the experiment the females of Wistar rats for 6 weeks after oophorectomy daily intragastrically administered enalapril in a dose of 0.5 mg/kg daily 1 time per day administered in addition drinkers to drink a mixture of solutions gomeopaticheskih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide person - C12, C30, C200.



 

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