Abstract
Aim:
To investigate the protective effects of simvastatin (Sim) combined with nifedipine (Nif) on endothelial cells and elucidate the action mechanism.
Methods:
Human umbilical vein endothelial cells (HUVEC) were used. mRNA and protein levels were measured by using reverse-transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively. Intracellular calcium and reactive oxygen species (ROS) were detected using confocal microscopy. The Griess assay was used to evaluate nitric oxide (NO) release.
Results:
Treatment of HUVEC with H2O2 100 μmol/L for 30 min inhibited the mRNA and protein expression of endothelial nitric oxide synthase (eNOS). With increased concentrations of Nif, eNOS mRNA and protein levels increased (P<0.05). Combined treatment with Sim 1.0 μmol/L and Nif 1.0 μmol/L significantly increased the mRNA and protein expression of eNOS and NO release compared with Sim or Nif alone (P<0.05). The combination significantly lowered the intracellular ROS level (P<0.05), which was correlated with the increase in eNOS and NO, but there was no visible change in intracellular calcium (P>0.05). Compared with individual drug treatment, Akt phosphorylation and the ratio of p-eNOS/eNOS were up-regulated in the combination group, and this effect was inhibited by the phosphatidylinositol 3-kinase (PI3K) inhibitors wortmannin and LY294002.
Conclusion:
The Sim-Nif combination effectively protects HUVEC against H2O2 injury by inhibiting intracellular ROS generation, increasing the ratio of p-eNOS/eNOS and up-regulating Akt phosphorylation.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Jiang JY, Wong MC, Zhang XH, Fung H, Griffiths S, Mercer S . Profiles of mortality among Chinese hypertensive patients in Hong Kong: a cohort study. J Hum Hypertens 2009; 23: 735–42.
Matthys KE, Bult H . Nitric oxide function in atherosclerosis. Mediators Inflamm 1997; 6: 3–21.
Ross R . The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature 1993; 362: 801–9.
Noll G, Tschudi M, Nava E, Luscher TF . Endothelium and high blood pressure. Int J Microcirc Clin Exp 1997; 17: 273–9.
Moncada S, Palmer RM, Higgs EA . Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 1991; 43: 109–42.
Rabelink TJ, Luscher TF . Endothelial nitric oxide synthase: host defense enzyme of the endothelium? Arterioscler Thromb Vasc Biol 2006; 26: 267–71.
Zanchetti A . The antiatherogenic effects of antihypertensive drugs: experimental and clinical evidence. Clin Exp Hypertens A 1992; 14: 307–31.
Lichtlen PR, Hugenholtz PG, Rafflenbeul W, Hecker H, Jost S, Deckers JW . Retardation of angiographic progression of coronary artery disease by nifedipine. Results of the International Nifedipine Trial on Antiatherosclerotic Therapy (INTACT). INTACT Group Investigators. Lancet 1990; 335: 1109–13.
Matsubara M, Hasegawa K . Benidipine, a dihydropyridine-calcium channel blocker, prevents lysophosphatidylcholine-induced injury and reactive oxygen species production in human aortic endothelial cells. Atherosclerosis 2005; 178: 57–66.
Deng HF, Xiong Y . Effect of pravastatin on impaired endothelium-dependent relaxation induced by lysophosphatidylcholine in rat aorta. Acta Pharmacol Sin 2005; 26: 92–8.
Ma FX, Liu LY, Xiong XM . Protective effects of lovastatin on vascular endothelium injured by low density lipoprotein. Acta Pharmacol Sin 2003; 24: 1027–32.
Gerson RJ, MacDonald JS, Alberts AW, Kornbrust DJ, Majka JA, Stubbs RJ, et al. Animal safety and toxicology of simvastatin and related hydroxy-methylglutaryl-coenzyme A reductase inhibitors. Am J Med 1989; 87: 28S–38S.
Todd PA, Goa KL . Simvastatin. A review of its pharmacological properties and therapeutic potential in hypercholesterolaemia. Drugs 1990; 40: 583–607.
Ye G, Metreveli NS, Ren J, Epstein PN . Metallothionein prevents diabetes-induced deficits in cardiomyocytes by inhibiting reactive oxygen species production. Diabetes 2003; 52: 777–83.
Brunt KR, Fenrich KK, Kiani G, Tse MY, Pang SC, Ward CA, et al. Protection of human vascular smooth muscle cells from H2O2-induced apoptosis through functional codependence between HO-1 and AKT. Arterioscler Thromb Vasc Biol 2006; 26: 2027–34.
Koneru S, Penumathsa SV, Thirunavukkarasu M, Samuel SM, Zhan L, Han Z, et al. Redox regulation of ischemic preconditioning is mediated by the differential activation of caveolins and their association with eNOS and GLUT-4. Am J Physiol Heart Circ Physiol 2007; 292: H2060–72.
Ignarro LJ, Buga GM, Wood KS, Byrns RE, Chaudhuri G . Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proc Natl Acad Sci USA 1987; 84: 9265–9.
Huang PL, Huang Z, Mashimo H, Bloch KD, Moskowitz MA, Bevan JA, et al. Hypertension in mice lacking the gene for endothelial nitric oxide synthase. Nature 1995; 377: 239–42.
Hyndman ME, Parsons HG, Verma S, Bridge PJ, Edworthy S, Jones C, et al. The T-786→C mutation in endothelial nitric oxide synthase is associated with hypertension. Hypertension 2002; 39: 919–22.
Mak IT, Boehme P, Weglicki WB . Protective effects of calcium channel blockers against free radical-impaired endothelial cell proliferation. Biochem Pharmacol 1995; 50: 1531–4.
Henrich D, Seebach C, Wilhelm K, Marzi I . High dosage of simvastatin reduces TNF-alpha-induced apoptosis of endothelial progenitor cells but fails to prevent apoptosis induced by IL-1beta in vitro. J Surg Res 2007; 142: 13–9.
Ungvari Z, Csiszar A, Kaley G . Vascular inflammation in aging. Herz 2004; 29: 733–40.
Hansson GK . Inflammation, atherosclerosis, and coronary artery disease. N Engl J Med 2005; 352: 1685–95.
Aono Y, Ariyoshi H, Sakon M, Ueda A, Tsuji Y, Kawasaki T, et al. Human umbilical vein endothelial cells (HUVECs) show Ca2+ mobilization as well as Ca2+ influx upon hypoxia. J Cell Biochem 2000; 78: 458–64.
Himmel HM, Whorton AR, Strauss HC . Intracellular calcium, currents, and stimulus-response coupling in endothelial cells. Hypertension 1993; 21: 112–27.
Cominacini L, Fratta Pasini A, Garbin U, Pastorino AM, Davoli A, Nava C, et al. Antioxidant activity of different dihydropyridines. Biochem Biophys Res Commun 2003; 302: 679–84.
Mason RP, Trumbore MW . Differential membrane interactions of calcium channel blockers. Implications for antioxidant activity. Biochem Pharmacol 1996; 51: 653–60.
Lenasi H, Kohlstedt K, Fichtlscherer B, Mulsch A, Busse R, Fleming I . Amlodipine activates the endothelial nitric oxide synthase by altering phosphorylation on Ser1177 and Thr495. Cardiovasc Res 2003; 59: 844–53.
Yamagishi S, Nakamura K, Takenaka K, Matsui T, Inoue H . Pleiotropic effects of nifedipine on atherosclerosis. Curr Pharm Des 2006; 12: 1543–7.
Yamagishi S, Nakamura K, Matsui T . Role of oxidative stress in the development of vascular injury and its therapeutic intervention by nifedipine. Curr Med Chem 2008; 15: 172–7.
Thomas SR, Witting PK, Drummond GR . Redox control of endothelial function and dysfunction: molecular mechanisms and therapeutic opportunities. Antioxid Redox Signal 2008; 10: 1713–65.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chen, Xn., Xu, J., Feng, Z. et al. Simvastatin combined with nifedipine enhances endothelial cell protection by inhibiting ROS generation and activating Akt phosphorylation. Acta Pharmacol Sin 31, 813–820 (2010). https://doi.org/10.1038/aps.2010.58
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/aps.2010.58
Keywords
This article is cited by
-
Fluvastatin attenuated the effect of expression of β1 integrin in PAN-treated podocytes by inhibiting reactive oxygen species
Molecular and Cellular Biochemistry (2015)
-
Current Therapeutic Strategies to Mitigate the eNOS Dysfunction in Ischaemic Stroke
Cellular and Molecular Neurobiology (2012)
