Modulation of Calcium Signaling of Angiotensin AT₁, Endothelin ET_A, and ET_B Receptors by Silibinin, Quercetin, Crocin, Diallyl Sulfides, and Ginsenoside Rb1

Abstract

Angiotensin II and endothelin-1 are potent vasoconstrictive peptides that play a central role in blood pressure regulation. Both peptides exert their pleiotropic effects via binding to their respective G-protein-coupled receptors, i.e., angiotensin AT₁ and endothelin type A and type B receptors. In the present study, we have selected six structurally different plant-derived compounds with known cardioprotective properties to evaluate their ability to modulate calcium signaling of the above-mentioned receptors. For this purpose, we used and validated a cellular luminescence-based read-out system in which we measured intracellular calcium signaling in Chinese hamster ovary cells that express the calcium sensitive apo-aequorin protein. Firstly, silibinin, a flavanolignan that occurs in milk thistle (Silybum marianum), was investigated and found to be an antagonist for the human angiotensin AT₁ receptor with an affinity constant of about 9 µM, while it had no effect on endothelin type A or type B receptor activation. Quercetin and crocin partially impeded intracellular calcium signaling resulting in a non-receptor-related reduction of the responses recorded for the three investigated G-protein-coupled receptors. Two organosulfur compounds, diallyl disulfide and diallyl trisulfide, as well as the triterpene saponin ginsenoside Rb1 did not affect the activation of the angiotensin AT₁ and endothelin type A and type B receptors. In conclusion, we were able, by using a nonradioactive cellular read-out system, to identify a novel pharmacological property of the flavanolignan silibinin.

Key words

angiotensin II - endothelin-1 - G-protein coupled receptors - silibinin - calcium signaling

• References

• 1 de Gasparo M, Catt KJ, Inagami T, Wright JW, Unger T. International union of pharmacology. XXIII. The angiotensin II receptors. Pharmacol Rev 2000; 52: 415-472
• 2 Atlas SA. The renin-angiotensin aldosterone system: pathophysiological role and pharmacologic inhibition. J Manag Care Pharm 2007; 13: 9-20
• 3 Weir MR, Dzau VJ. The renin-angiotensin-aldosterone system: a specific target for hypertension management. Am J Hypertens 1999; 12: 205S-213S
• 4 Miyauchi T, Masaki T. Pathophysiology of endothelin in the cardiovascular system. Annu Rev Physiol 1999; 61: 391-415
• 5 Nakas-Icindic E, Zaciragic A, Hadzovic A, Avdagic N. Endothelin in health and disease. Bosn J Basic Med Sci 2004; 4: 31-34
• 6 Davenport AP. International Union of Pharmacology. XXIX. Update on endothelin receptor nomenclature. Pharmacol Rev 2002; 54: 219-226
• 7 Davenport AP, Battistini B. Classification of endothelin receptors and antagonists in clinical development. Clin Sci (Lond) 2002; 103 (Suppl. 48) 1S-3S
• 8 Maione F, Cicala C, Musciacco G, De Feo V, Amat AG, Ialenti A, Mascolo N. Phenols, alkaloids and terpenes from medicinal plants with antihypertensive and vasorelaxant activities. A review of natural products as leads to potential therapeutic agents. Nat Prod Commun 2013; 8: 539-544
• 9 Stoclet JC, Chataigneau T, Ndiaye M, Oak MH, El Bedoui J, Chataigneau M, Schini-Kerth VB. Vascular protection by dietary polyphenols. Eur J Pharmacol 2004; 500: 299-313
• 10 Ulrich-Merzenich G, Panek D, Zeitler H, Vetter H, Wagner H. Drug development from natural products: exploiting synergistic effects. Indian J Exp Biol 2010; 48: 208-219
• 11 Gazak R, Walterova D, Kren V. Silybin and silymarin–new and emerging applications in medicine. Curr Med Chem 2007; 14: 315-338
• 12 Li Volti G, Salomone S, Sorrenti V, Mangiameli A, Urso V, Siarkos I, Galvano F, Salamone F. Effect of silibinin on endothelial dysfunction and ADMA levels in obese diabetic mice. Cardiovasc Diabetol 2011; 10: 62
• 13 Ai W, Zhang Y, Tang QZ, Yan L, Bian ZY, Liu C, Huang H, Bai X, Yin L, Li H. Silibinin attenuates cardiac hypertrophy and fibrosis through blocking EGFR-dependent signaling. J Cell Biochem 2010; 110: 1111-1122
• 14 Khurana S, Venkataraman K, Hollingsworth A, Piche M, Tai TC. Polyphenols: benefits to the cardiovascular system in health and in aging. Nutrients 2013; 5: 3779-3827
• 15 Imenshahidi M, Razavi BM, Faal A, Gholampoor A, Mousavi SM, Hosseinzadeh H. Effects of chronic crocin treatment on desoxycorticosterone acetate (doca)-salt hypertensive rats. Iran J Basic Med Sci 2014; 17: 9-13
• 16 Imenshahidi M, Razavi BM, Faal A, Gholampoor A, Mousavi SM, Hosseinzadeh H. The Effect of Chronic Administration of Saffron (Crocus sativus) Stigma Aqueous Extract on Systolic Blood Pressure in Rats. Jundishapur J Nat Pharm Prod 2013; 8: 175-179
• 17 Taira S, Ikeda R, Yokota N, Osaka I, Sakamoto M, Kato M, Sahashi Y. Mass spectrometric imaging of ginsenosides localization in Panax ginseng root. Am J Chin Med 2010; 38: 485-493
• 18 Wang CZ, Kim KE, Du GJ, Qi LW, Wen XD, Li P, Bauer BA, Bissonnette MB, Musch MW, Chang EB, Yuan CS. Ultra-performance liquid chromatography and time-of-flight mass spectrometry analysis of ginsenoside metabolites in human plasma. Am J Chin Med 2011; 39: 1161-1171
• 19 Li J, Shao ZH, Xie JT, Wang CZ, Ramachandran S, Yin JJ, Aung H, Li CQ, Qin G, Vanden Hoek T, Yuan CS. The effects of ginsenoside Rb1 on JNK in oxidative injury in cardiomyocytes. Arch Pharm Res 2012; 35: 1259-1267
• 20 Yun HM, Ban JO, Park KR, Lee CK, Jeong HS, Han SB, Hong JT. Potential therapeutic effects of functionally active compounds isolated from garlic. Pharmacol Ther 2014; 142: 183-195
• 21 Benavides GA, Squadrito GL, Mills RW, Patel HD, Isbell TS, Patel RP, Darley-Usmar VM, Doeller JE, Kraus DW. Hydrogen sulfide mediates the vasoactivity of garlic. Proc Natl Acad Sci U S A 2007; 104: 17977-17982
• 22 Nikolaou A, Van den Eynde I, Tourwe D, Vauquelin G, Toth G, Mallareddy JR, Poglitsch M, Van Ginderachter JA, Vanderheyden PM. [3H]IVDE77, a novel radioligand with high affinity and selectivity for the insulin-regulated aminopeptidase. Eur J Pharmacol 2013; 702: 93-102
• 23 Verheijen I, Vanderheyden PM, De Backer JP, Vauquelin G. AT1 receptor antagonists. Curr Med Chem Cardiovasc Hematol Agents 2004; 2: 69-77
• 24 Vanderheyden PM, Fierens FL, Vauquelin G. Angiotensin II type 1 receptor antagonists. Why do some of them produce insurmountable inhibition?. Biochem Pharmacol 2000; 60: 1557-1563
• 25 Arunlakshana O, Schild HO. Some quantitative uses of drug antagonists. Br J Pharmacol Chemother 1959; 14: 48-58
• 26 Caballero-George C, Vanderheyden PM, De Bruyne T, Shahat AA, Van den Heuvel H, Solis PN, Gupta MP, Claeys M, Pieters L, Vauquelin G, Vlietinck AJ. In vitro inhibition of [3H]-angiotensin II binding on the human AT1 receptor by proanthocyanidins from Guazuma ulmifolia bark. Planta Med 2002; 68: 1066-1071
• 27 Singh RP, Tyagi AK, Zhao J, Agarwal R. Silymarin inhibits growth and causes regression of established skin tumors in SENCAR mice via modulation of mitogen-activated protein kinases and induction of apoptosis. Carcinogenesis 2002; 23: 499-510
• 28 Zhou B, Wu LJ, Li LH, Tashiro S, Onodera S, Uchiumi F, Ikejima T. Silibinin protects against isoproterenol-induced rat cardiac myocyte injury through mitochondrial pathway after up-regulation of SIRT1. J Pharmacol Sci 2006; 102: 387-395
• 29 Dayoub O, Andriantsitohaina R, Clere N. Pleiotropic beneficial effects of epigallocatechin gallate, quercetin and delphinidin on cardiovascular diseases associated with endothelial dysfunction. Cardiovasc Hematol Agents Med Chem 2013; 11: 249-264
• 30 Duarte J, Perez Vizcaino F, Utrilla P, Jimenez J, Tamargo J, Zarzuelo A. Vasodilatory effects of flavonoids in rat aortic smooth muscle. Structure-activity relationships. Gen Pharmacol 1993; 24: 857-862
• 31 Perez-Vizcaino F, Ibarra M, Cogolludo AL, Duarte J, Zaragoza-Arnaez F, Moreno L, Lopez-Lopez G, Tamargo J. Endothelium-independent vasodilator effects of the flavonoid quercetin and its methylated metabolites in rat conductance and resistance arteries. J Pharmacol Exp Ther 2002; 302: 66-72
• 32 Perez A, Gonzalez-Manzano S, Jimenez R, Perez-Abud R, Haro JM, Osuna A, Santos-Buelga C, Duarte J, Perez-Vizcaino F. The flavonoid quercetin induces acute vasodilator effects in healthy volunteers: correlation with beta-glucuronidase activity. Pharmacol Res 2014; 89: 11-18
• 33 Perez-Vizcaino F, Duarte J. Flavonols and cardiovascular disease. Mol Aspects Med 2010; 31: 478-494
• 34 Khoo NK, White CR, Pozzo-Miller L, Zhou F, Constance C, Inoue T, Patel RP, Parks DA. Dietary flavonoid quercetin stimulates vasorelaxation in aortic vessels. Free Radic Biol Med 2010; 49: 339-347
• 35 Nifli AP, Theodoropoulos PA, Munier S, Castagnino C, Roussakis E, Katerinopoulos HE, Vercauteren J, Castanas E. Quercetin exhibits a specific fluorescence in cellular milieu: a valuable tool for the study of its intracellular distribution. J Agric Food Chem 2007; 55: 2873-2878
• 36 Boskabady MH, Aslani MR. Relaxant effect of Crocus sativus (saffron) on guinea-pig tracheal chains and its possible mechanisms. J Pharm Pharmacol 2006; 58: 1385-1390
• 37 He SY, Qian ZY, Tang FT. Effect of crocin on intracellular calcium concentration in cultured bovine aortic smooth muscle cells. Yao Xue Xue Bao 2004; 39: 778-781
• 38 Demaegdt H, Vanderheyden P, De Backer JP, Mosselmans S, Laeremans H, Le MT, Kersemans V, Michotte Y, Vauquelin G. Endogenous cystinyl aminopeptidase in Chinese hamster ovary cells: characterization by [125I]Ang IV binding and catalytic activity. Biochem Pharmacol 2004; 68: 885-892
• 39 Le Poul E, Hisada S, Mizuguchi Y, Dupriez VJ, Burgeon E, Detheux M. Adaptation of aequorin functional assay to high throughput screening. J Biomol Screen 2002; 7: 57-65
• 40 Iredale PA, Hill SJ. Increases in intracellular calcium via activation of an endogenous P2-purinoceptor in cultured CHO-K1 cells. Br J Pharmacol 1993; 110: 1305-1310
• 41 Lecoeur H, Ledru E, Prevost MC, Gougeon ML. Strategies for phenotyping apoptotic peripheral human lymphocytes comparing ISNT, annexin-V and 7-AAD cytofluorometric staining methods. J Immunol Methods 1997; 209: 111-123