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

FIELD: medicine.

SUBSTANCE: 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 lozartan 6 mg/kg and daily introduction of mixed solutions of homoeopathic dilutions of polyclonal rabbit C12, C30, C200 antibodies to human endothelial nitrogen oxide synthase once a day added to drinking bowls.

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 hypoestrogenic conditions.

1 ex, 2 tbl

 

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, RF patent No. 2329807.

The main disadvantage of this method is that the correction geoastro-meningocerebral 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 is 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 losartan with endothelial dysfunction due is an over stimulation AT 2 subtype angiotensin II receptor and implemented due bradykininase ways of endotheliopathy, 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 antagonists of the angiotensin II receptor (AT 1-subtype) and the mixture 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 losartan at a dose of 6 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 the province of the Dili with bilateral oophorectomy. 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 100 MP, production Biopac System, Inc., USA. Functional tests: endothelium-dependent vasodilatation (ESV) - intravenous administration of acetylcholine (ach) at a dose of 40 mcg/kg, endothelium-independent vasodilatation (ANSW) - intravenous 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.

An example of a specific implementation.

Bolus intravenous AH 3-5 with led to recompeting 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 with 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 was 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 that have this indicator SOS is avsl 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 losartan 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 + losartanSource140,1±7,0**107,5±5,2**405,0±14,6
OHand 82.2±3,744,2±4,5397,8±13,8
NPfor 96.1±5,857,7 is a 6.2 401,3±6,8
Hypoestrogenic state of + losartan + blend solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial NOS man - C12, C30, C200Source135,1 to±6.5**98,7±8,7**382,6±9,9
ESV with AH94,0±5,3*53,9±6,9*379,4±10,1
ANSV with NP106,7±5,5**57,7±6,0*to 369.6±12,9
* - p<0,05 in comparison with the group intact; ** - p<0,05 in comparison with the group with hypoestrogenism state

Intragastric administration of losartan at a dose of 6 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) led to the decline of the initial values of AD (GARDEN - 135,1±6,5, DBP - 98,7±8.7 mm Hg) (table 1).

Also in this group, where the background of bilateral oophorectomy for 6 weeks was administered losartan at a dose of 6 mg/kg and the mixture solutions homeopathy esih dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide person - C12, C30, C200 QED was 1.1, which was significantly lower than in the group with hypoestrogenism condition. Whereas, in the group with losartan monotherapy, this figure amounted to 1.5 (table 2).

Table 2
Indicators that reflect the correction hypoestrogen-induced endothelial dysfunction in the background of bilateral oophorectomy with losartan 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 time of the vascular response (s)The area of the vascular response (used)QED, usled
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 + losartanOH61,5±4,442,6±3,6**1323,6±166,71,5±0,2**
NP51,8±5,0**34,9±3,0**1984,5±247,2
Hypoestrogenic state of + losartan + blend solutions of homeopathic dilutions of polyclonal antibodies to endothelial NOS man - C12, C30, C200OH43,6±6,266,6±6,11349,8±105,11,1±0,1**
NP39,6±4,6of 77.0±7,41440,6±109,7
* - p<0,05 in comparison with the group intact, ** - p<0,05 in comparison with grupoj with hypoestrogenism state

Thus, the obtained results allow to conclude the correction hypoestrogen-induced endothelial amid bilateral oophorectomy with losartan 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 losartan at a dose of 6 mg/kg daily 1 time per day administered in addition drinkers to drink a mixture of solutions of homeopathic dilutions of polyclonal rabbit antibodies to endothelial synthase nitric oxide person - C12, C30, C200.



 

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