The article presents the results of the additive endothelio- and cardioprotective effects of the combination of L-

Transcription

1 ISSN: X CODEN: IJPTFI Available Online through Research Article COMBINED PHARMACOLOGICAL CORRECTION OF THE METABOLIC PATHWAYS OF L-ARGININE/NOHYPOESTROGENEMIA IN THE SIMULATION OF DEFICIENCY OF NITRIC OXIDE Tatiana G.Pokrovskaya*, Michail V.Korokin, Oleg S. Gudyrev,Lev N.Sernov,Ol'ga A.Osipova, Taisia A.Chadieva, Anton P. Dovgan Belgorod State Univercity, Russia, , Belgorod, Pobedyst Received on Accepted on Abstract The article presents the results of the additive endothelio- and cardioprotective effects of the combination of L- arginine with the products of the major antihypertensive groups enalapril, losartan, amlodipine, indapamide, nebivolol on the model of hypoestrogen-induced endothelial dysfunction, which allows us to recommend these combinations for clinical study.additive effect is maximal where the point of application of the pharmacodynamic action of L-arg as a precursor of NO and the competitive ADMA-inhibitor, is different from the mechanism of endothelioprotection by known antihypertensive drugs in ovariectomy. Keywords: Ovariectomy, L-arginine, enalapril, losartan, amlodipine, indapamide, nebivolol. Introduction There are several groups of factors may play a role of increasethe risk of cardiovascular disease in menopause. Such as disfunction of endothelial cells, changes in heart function, metabolic activation and release of asymmetric dimethylarginine (ADMA) an endogenous inhibitor of NO-synthase. [1-4]. Currently one of the most feasible methods of increasing the level of nitric oxide (NO) and overcoming the blocking action of ADMA is the injection into the body of L-arginine [5-13]. The aim of the present study was to investigate the effects of L-arginine, enalapril, losartan, indapamid and nebivolol and their combinations on the performance of the endothelium athypoestrogen-induced endothelial dysfunction (ED) model. Methods The experiments were made on 120 adult female rats of Wistar weighing g, which under anesthesia (Zoletil 50 mg/kg) underwent bilateral ovariectomy (OE) (n=10). Animals of intact group (IA) (n=10) were exposed to false IJPT Sep-2016 Vol. 8 Issue No Page 15175

2 operation without removal of ovaries [14-16].Enalapril maleate (E), losartan potassium (L), amlodipine (A), indapamid (I), nebivolol (N) used in doses of 0.5 mg/kg; 6 mg/kg; 0.5 mg/kg; 2 mg/kg; 0.5 mg/kg, respectively and injected intragastricaly one time a day. L-arginine (L-arg) was injected at the dose of 200 mg/kg once a day intraperitoneally. Investigational drugs and their combination with L-arginine were injected simultaneously in the same time within 42 days after the OE [17, 18]. IA were injected saline in the same volume (n=10). On the 43 rd day from the beginning of the experiment under anesthesia (Zoletil 50 mg/kg) were catheterized the left carotid artery and the cavity of the left ventricle of the heart to the registration of hemodynamic parameters (systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), left ventricular pressure (LVP)) with the help of the sensor TSD104A and hardware-software complex MP100(Biopac System, Inc., USA) bolus into the right femoral vein was injected acetylcholine (40 μg/kg) an endothelium dependent vasodilation - EDVD, nitroprusside sodium (30 mg/kg) endothelium-independent dilatation (EIVD), adrenaline hydrochloride 1ˑ10-5 mol/l adrenoreactivity (ADR). The test with load of the left ventricular resistance was carried out (clamping the ascending aorta for 30 sec). The definition of stable NO metabolites (NOx) was carried out one-step method for quantitative determination of total nitrate and nitrite [19]. Determination of cardiomyocyte diameter (DM) was carried by the method of G. G. Avtandilov [20]. Results and Discussion The ratio of the area EIND to EDVD in terms of the average blood pressure is mathematically presented in the form of coefficient of endothelial dysfunction (CED) [21, 22, 23]. CED in the IA group was 1.1±0.09, and afteroe 2.1±0.2, the values of SBP/DBP 160.0±6.2/124.9±5.5 mm Hg. The OE has led to an increase the LVP in sample into ADR and reduction of myocardial reserve in the sample with the load resistance (LR). So, IA LVP to 25th second clamping the aorta (LVP 25 ) decreased to 89.3±4.2%, and in animals with deficiency of estrogen to 67.9±2.1%. The concentration of nitrite ions (NO x ) in the OE 71.5±4.1, whereas IA 122.8±4.5 µmol. Our studies have shown that L-arginine has a strong endothelioprotective effect, comparable with the other study drugs.ced in group OE+Larg was 1.3±0.1, and OE+E is 1.3±0.1, and OE+L 1.3±0.1; OE+A 1.5±0.09; OE+N 1.1±0.1; OE+I 1.4±0.1. Only in the case of therapy N the development of hypertension was prevent, has a moderate hypotensive effect of other drugs, except I. With the injection of the drugs under study registered a decline of LVP during the tests on ADR: IA ± 7.1 and OE ±6.4, the greatest decrease of myocardial contractility was noted within the injection of L-arg (210.9±9.4), N (200.5±8.7), I (210.4±7.6 mm Hg), the other drugs occupy an intermediate position. IJPT Sep-2016 Vol. 8 Issue No Page 15176

3 A similar trend identified in the sample by LR efficient were all studied substances, and a little less E and L. The concentration of nitrite ions in blood during monotherapy was significantly increased only with the introduction of L- arg and N (107.9±15.7 and 99.1±12.1 μmol, respectively). Morphometric studies showed that all the studied substances in different degree prevent the development of miocardiocytes, this effect is expressed in the largest extent at L-arg, L and N. Thus, by the combination of functional, biochemical and morphological indicators of endothelioprotection L-arg showed the effects of activity comparable or superior to that of drugs with proved endothelioprotective activity. Table 1: The effect of combined use of L-arginine with the products of major antihypertensive drugs on blood pressure (SBP, DBP), the coefficient of endothelial dysfunction (CED) and adrenoreactivity (ADR) in the simulation of hypoestrogen-induced deficit (OE) of the nitric oxide (M±m). ADR SBP, DBP, CED, Groups of animals (LVP, mm mm Hg mm Hg c.u. Hg) IA ± ± ± ± 6.3 OE ± 6.2* ± 5.5* 2.1±0.2* 243.3±6.4* OE+L-arginine ± ± ± 0.9±0.1** 9.7** 5.7** 12.5** OE+enalapril 139.6±7.3** 107.4±4.9** 1.3± ± 0.1** 11.3** OE + losartan 140.1±7.0** 107.5±5.2** 1.5±0.2** ± 10.1** OE + amlodopine 154.1± ± ±0.2** ± 9.3** OE + nebivolol 125.1±4.8** 94.8±4.1** 1.1±0.1** 200.5±6.4** OE + indapamid 156.4± ± ±0.4** 210.4±5.3** ± OE +enalapril +L-arginine ±6.9** у 0.8±0.1** 205.3±2.4** y 4.5** 130.4±5.7** OE + losartan + L-arginine у 98.0±4.1** у 0.8±0.2** 210.7±6.1** OE + amlodipine +L-arginine 134.2±7.8** у 101.0±5.7** у 0.6±0.1** 182.2±16.4** y OE + nebivolol +L-arginine 121.1±3.1** 90.2±3.1** 0.9± ±1.0** OE + indapamid +L-arginine 137.2±5.1** у 98.1±3.4** у 1.1±0.1** 206.0±3.9** Note: SBP, DBP systolic and diastolic blood pressure, LVP left ventricular pressure, * - p<0.05 compared with intact rats; **- p<0.05 compared with the OE group; y p<0.05 compared with the group receiving monotherapy with the drug in the appropriate dose. The study endothelioprotective activity of combined application of L-argwith known antihypertensive drugs with the OE showed that the values of CED in comparison with monotherapy is more pronounced closer to the values of IA. Digital expression of this index in the studied IJPT Sep-2016 Vol. 8 Issue No Page 15177

4 groups was L-arg + E 0.8±0.1; L-arg + L 0.8±0.09; L-arg + A 0.6±0.08; L-arg + N 0.9±0.09; L-arg + I 1.1±0.1. Additional hypotensive effect in the OE was observed in almost all combinations, to a lesser extent in the combinations L-arg +A and L-arg + I (table 1). A sample of ADR revealed a further reduction in LVP in all groups of animals received L-arg in combination with the study drugs, but to a lesser degree with losartan. Table 2. The effect of combined use of L-arginine with the major antihypertensive drugs on the myocardial reserve exhaustion during the tests on the load resistance (LR), the concentration of nitrite ions (NO x ) and the diameter of cardiomyocytes (DM) in model ofhypoestrogen-induced deficit (OE) of the nitric oxide (M±m). Groups of animals LVP 25, NO x, DM, % μmol μm IA 88.7± ± ±0.2 OE 67.9±2.1* 75.1±13.5* 12.5±0.7* OE+L-arginine 87.2±3.7** 98.6±7.7** 9.2±0.1** OE+enalapril 81.6±4.3** 65.3± ±0.1** OE + losartan 74.9±5.0** 71.1± ±0.2** OE + amlodopin 86.7±3.7** 68.1± ±0.2** OE + nebivolol 89.9±4.9** 101.2±4.1** 9.6±0.1** OE + indapamid 83.6±3.6** 63.2± ±0.4** OE +enalapril +L-arginine 88.7±4.9** у 113.1±12.5** у 8.9±0.1** OE + losartan + L-arginine 81.5±3.7** у 109.5±4.1** у 8.7±0.2** OE + amlodipine +L-arginine 89.5±4.8** 101.7±5.7** у 9.3±0.1** OE + nebivolol +L-arginine 92.2±5.1** 117.6±3.1** 8.8±0.2 OE + indapamid +L-arginine 90.6±4.1** у 99.2±3.4** у 9.1±0.1** Note:SBP, DBP systolic and diastolic blood pressure; LVP 25 is the ratio of the increment values on LVP at 25 seconds to LVP at 5 seconds after cross-clamping of the aorta expressed in %; * - at p<0.05 compared with intact rats; **- p<0.05 compared with the OE group; y p<0.05 compared with the group receiving monotherapy with the drug in the appropriate dose. LR revealed a significant restoration of myocardial reserve compared with monotherapy in all the groups studied in combination with L-arg, the best growth was observed in groups E and L (85.8±6.9% and 81.5±6.1% respectively) (Fig. 2 LR ). Maximal preservation of myocardial reserve was observed in group L-arg + N. NO x in the combined use was significantly higher than in the appropriate monotherapy, approaching the values in IA, most in the group L-arg + A and L-arg + I. The combinations of L-arg with the studied drugs are almost completely prevented hypertrophy of myocardiocytes (table 2). Integral planimetric analysis of the obtained data (SBP, DBP, CED, LVP 25, NO x ) to assess the degree of additivity of endothelio - and cardioprotective effects of L-arg was IJPT Sep-2016 Vol. 8 Issue No Page 15178

5 performed using method of comparing of the space vector diagrams constructed with the help of named indicators, in groups with the injection of drugs in relation to group IA and OE. We calculated coefficient of addition addition of the difference between the size of space under the resulting vector diagram of each investigational drug and the area of its combination with L-arg. L-arg for all combinations encreaseendothelio - and cardioprotective activity against hypoestrogen-induced deficit of nitrogen oxide [24,25]. Additive effect is maximal where the point of application of the pharmacodynamic action of L-arg as a precursor of NO and the competitive ADMA-inhibitor, is different from the mechanism of endothelioprotection by known antihypertensive drugs in OE. Conclusion L-arginine showed the additive endotelio and cardiotropic effects in combination with known antihypertensive drugs in the correction of cardiovascular complications in hypoestrogen-induced endothelial disfunction. In ascending values of the coefficient of additio, studied combinations are as follows: L-arginine + nebivolol< L-arginine + amlodipine< L-arginine + indapamid< L-arginine + losartan < L-arginine + amlodipin. References 1. Karkanaki, A., et al., Hormone therapy and asymmetrical dimethylarginine in postmenopausal women. Hormone, 9(2): Kocak, H., et al., Serum asymmetric dimethylarginine and nitric oxide levels in obese postmenopausal women, J Clin Lab Anal., 25(3): Vladimirova-Kitova, L.G., Asymmetric dimethylarginine-mechanisms and targets for therapeutic management.folia Med (Plovdiv), 50(1): Wang, J., Aim,S., Wang,Х.and D.Wilcken, L-arginine regulates asymmetric dimethylarginine metabolism by inhibiting dimethylargininedimethylaminohydrolase activity in hepatic (HepG2) cells. Cell Mol Life Sci., 63(23): Böger, G., Rudolph,T., Maas,R. et al., 2007.Asymmetric dimethylarginine determines the improvement of endothelium-dependent vasodilation by simvastatin: Effect of combination with oral L-arginine. J Am CollCardiol, 49(23): Pokrovskii, M., Kochkarov,V., Pokrovskaya,T.et al., Comparative study of potential endothelioprotectors and impaza in modeled nitric oxide deficiency. Bulletin of Experimental Biology and Medicine, 148(3): (In Russia). IJPT Sep-2016 Vol. 8 Issue No Page 15179

6 7. Tyurenkov, I., Voronkov,A. and A. Robertus, The Study of Endothelial Dysfunction in Animals With Experimentally Induced Deficiency of Sex Hormones. Cardiology, 49(5): (In Russia). 8. Pokrovskaya,T., Pokrovskii,M., Kochkarov,V. et al., 2006.The study endotelioprotective effect of L-arginine and its combination with enalapril and losartan. Biomedicine, 1(4): (In Russia). 9. Pokrovskii, M., Pokrovskaya,T., Gureev,V. et al., Correction of endothelial dysfunction by L-arginine under experimental pre-eclampsia conditions. Eksperimental'nayaiKlinicheskayaFarmakologiya, 75 (2): 14-16(In Russia). 10. Korokin, M., Pokrovskii,M., Gudyrev,O.et al., Pharmacological correction of endothelial dysfunction in rats using e-nos cofactors. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 6 (5): Denisyuk, T., Lazareva,G., Provotorov,V.and A. Shaposhnikov, Endothelium and cardioprotective effects of HMG-Co-A-reductase in combination with L-arginine in endothelial dysfunction modeling. Research result: pharmacology and clinical pharmacology, 2(1): Shakhno, E., Savitskaya,T., Pokrovskaya,T., Yakushev,V., Pokrovsky,M. and D. Grinshpan, Use of L- arginine immobilised on activated carbon for pharmacological correction of endothelial disfunction. Research result: pharmacology and clinical pharmacology, 2(1): Korokin, M., Pokrovskii,M., Kochkarov,V.et al., Endothelial and cardio protective effects of tetrahydrobiopterin, L-norvaline, L-arginine and their combinations by simulation of hyperhomo-cysteine induced endothelial dysfunction. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 5 (6): Kochkarov, V., Molchanova, O., Pokrovskii, M. et al., Endothelium-protective action of thioctic acid and rosuvastatin combination at concomitant hypoestrogen and L-Name-induced deficit of nitric oxide. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 5(5): Rajkumar, D., Faitelson,A., Gudyrev, O.et al., Comparative evaluation of enalapril and losartan in pharmacological correction of experimental osteoporosis and fractures of its background. Journal of Osteoporosis. Date Views http:// (In Russia). IJPT Sep-2016 Vol. 8 Issue No Page 15180

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