Synergistic Effect of Amlodipine and Atorvastatin in Reversing LDL-Induced Endothelial Dysfunction
Statins and certain calcium channel blockers may improve nitric oxide (NO) release and endothelial function through various mechanisms, but their combined effects are not well understood.
The separate versus combined effects of amlodipine (AML) and atorvastatin (AT) on NO and peroxynitrite (ONOO−) were measured in human umbilical vein endothelial cells (HUVEC) in the presence and absence of low-density lipoprotein (LDL) using electrochemical nanosensors.
The combination of AML (5 μmol/l) and AT (3-6 μmol/l) directly stimulated NO release that was about twofold greater than the sum of their separate effects (p < 0.05). This synergistic activity is attributed to enhanced endothelial NO synthase (eNOS) function and decreased cytotoxic ONOO−. LDL (100 mg/dl) caused a dysfunction of HUVEC manifested by a 60% reduction in NO and an almost twofold increase in ONOO−. Treatment with AML/AT partially reversed the effects of LDL on endothelial function, including a 90% increase in NO and 50% reduction in ONOO−. Small-angle X-ray diffraction analysis indicates that AML and AT are lipophilic and share an overlapping molecular location in the cell membrane that could facilitate electron transfer for antioxidant mechanisms.
These findings indicate a synergistic effect of AML and AT on an increase in NO concentration, reduction of nitroxidative stress. Also, AML/AT partially restored the NO level of LDL-induced dysfunctional endothelium. Their combined effects may be enhanced by antioxidant properties related to their intermolecular actions in the cell membrane and an increase in the expression and coupling of endothelial nitric oxide synthase.
Key Wordsendothelium LDL nitric oxide oxidative stress
- 2.D. G. Harrison, P. C. Freiman, M. L. Armstrong, M. L. Marcus, and D. D. Heistad. Alterations of vascular reactivity in atherosclerosis. Circ. Res. 61:74–80 (1987).Google Scholar
- 11.J. D. Neaton and D. Wentworth. Serum cholesterol, blood pressure, cigarette smoking, and death from coronary heart disease. Overall findings and differences by age for 316,099 white men. Multiple Risk Factor Intervention Trial Research Group. Arch. Intern. Med. 152:56–64 (1992).PubMedCrossRefGoogle Scholar
- 17.L. Vergnani, S. Hatrik, F. Ricci, A. Passaro, N. Manzoli, G. Zuliani, V. Brovkovych, R. Fellin, and T. Malinski. Effect of native and oxidized low-density lipoprotein on endothelial nitric oxide and superoxide production: key role of l-arginine availability. Circulation 101(11):1261–1266 (2000).PubMedGoogle Scholar
- 22.G. B. Mancini, G. C. Henry, C. Macaya, B. J. O'Neill, A. L. Pucillo, R. G. Carere, T. J. Wargovich, H. Mudra, T. F. Luscher, M. I. Klibaner, H. E. Haber, A. C. Uprichard, C. J. Pepine, and B. Pitt. Angiotensin-converting enzyme inhibition with quinapril improves endothelial vasomotor dysfunction in patients with coronary artery disease. The TREND (Trial on Reversing Endothelial Dysfunction) Study. Circulation 94(3):240–243 (1996).Google Scholar
- 29.S. E. Nissen, E. M. Tuzcu, P. Libby, P. D. Thompson, M. Ghali, D. Garza, L. Berman, H. Shi, E. Buebendorf, and E. J. Topol. Effect of antihypertensive agents on cardiovascular events in patients with coronary disease and normal blood pressure: the CAMELOT study: a randomized controlled trial. JAMA 292:2217–2226 (2004).PubMedCrossRefGoogle Scholar
- 30.P. S. Sever, B. Dahlof, and N. R. Poulter. Prevention of coronary and stroke events with atorvastatin in hypertensive patients who have average or lower-than-average cholesterol concentrations, in the Anglo-Scandinavian Cardiac Outcomes Trial-Lipid Lowering Arm (ASCOT-LLA): a multicentre randomised controlled trial. Lancet 361:1149–1158 (2003).PubMedCrossRefGoogle Scholar
- 31.H. M. Colhoun, D. J. Betteridge, P. N. Durrington, G. A. Hitman, Neil HAW, S. J. Livingstone, M. J. Thomason, M. I. Mackness, V. Charlton-Menys, and J. H. Fuller. Primary prevention of cardiovascular disease with atorvastatin in type 2 diabetes in the Collaborative Atorvastatin Diabetes Study (CARDS): Multicentre randomised placebo-controlled trial. Lancet 364:685–696 (2004).PubMedCrossRefGoogle Scholar
- 32.P. Sever, B. Dahlof, N. Poulter, H. Wedel, G. Beevers, M. Caulfield, R. Collins, S. Kjeldsen, A. Kristinsson, G. McInnes, J. Mehlsen, M. Niemenem, E. O'Brien, and J. Ostergren. Potential synergy between lipid-lowering and blood-pressure-lowering in the Anglo-Scadinavian Cardiac Outcomes Trail. Eur. Heart J. 27:2982–2988 (2006).PubMedCrossRefGoogle Scholar
- 33.D. J. M. Delsing, J. W. Jukema, M. A. van de Wiel, J. J. Emeis, A. van der Laarse, L. M. Havekes, and H. M. G. Princen. Differential effects of amlodipine and atorvastatin treatment and their combination on atherosclerosis in ApoE*3-Leiden transgenic mice. J. Cardiovasc. Pharmacol. 42:63–70 (2003).PubMedCrossRefGoogle Scholar
- 38.P. Vallance, S. Patton, K. Bhagat, R. MacAllister, M. Radomski, S. Moncada, and T. Malinski. Direct measurement of nitric oxide in human beings. Anal. Chem. 346:153–154 (1995).Google Scholar
- 43.R. P. Mason and R. F. Jacob. X-ray diffraction analysis of membrane structure changes with oxidative stress. In D. Armstrong (ed.), Methods in Molecular Biology: Ultrastructural and Molecular Biology Protocols. Vol 193. Humana Press Inc., Totowa, NJ, 2002, pp. 71–80.Google Scholar
- 45.L. G. Herbette, T. MacAlister, T. F. Ashavaid, and R. A. Colvin. Structure-function studies of canine cardiac sarcolemmal membranes. II. Structural organization of the sarcolemmal membrane as determined by electron microscopy and lamellar X-ray diffraction. Biochim. Biophys. Acta. 812(3):609–623 (1985).PubMedCrossRefGoogle Scholar
- 46.K. K. Koh, M. J. Quon, S. H. Han, W. J. Chung, J. Y. Ahn, Y. H. Seo, M. H. Kang, T. H. Ahn, I. S. Choi, and E. K. Shin. Additive beneficial effects of losartan combined with simvastatin in the treatment of hypercholesterolemic, hypertensive patients. Circulation 110:3687–3692 (2004).PubMedCrossRefGoogle Scholar
- 49.D. W. Stepp, J. Ou, A. W. Ackerman, S. Welak, D. Klick, K. A. Pritchard Jr. Native LDL and minimally oxidized LDL differentially regulate superoxide anion in vascular endothelium in situ. Am J Physiol, Heart Circ Physiol. 283(2):H750–H759 (2002).Google Scholar
- 51.J. Martinez-Gonzalez, B. Raposo, C. Rodriguez, and L. Badimon. 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibition prevents endothelial NO synthase downregulation by atherogenic levels of native LDLs: balance between transcriptional and posttranscriptional regulation. Arterioscler. Thromb. Vasc. Biol. 21(5):804–809 (2001).PubMedGoogle Scholar