Phosphorylation of Endothelial NOS Contributes to Simvastatin Protection Against Myocardial No-reflow and Infarction in Reperfused Swine Hearts: Partially Via the PKA Signaling Pathway

Overview

Journal Acta Pharmacol Sin

Specialty Pharmacology

Date 2012 Jun 5

PMID 22659627

Citations 15

Authors

Xiang-Dong Li

Yue-Jin Yang

Yong-Jian Geng

Jing-Lin Zhao

Hai-Tao Zhang

Yu-Tong Cheng

Yi-Ling Wu

Affiliations

Soon will be listed here.

Abstract

Aim: The cholesterol-lowering drugs statins could enhance the activities of endothelial nitric oxide synthase (eNOS) and protect myocardium during ischemia and reperfusion. The aim of this study was to examine whether protein kinase A (PKA) was involved in statin-mediated eNOS phosphorylation and cardioprotection.

Methods: 6-Month-old Chinese minipigs (20-30 kg) underwent a 1.5-h occlusion and 3-h reperfusion of the left anterior descending coronary artery (LAD). In the sham group, the LAD was encircled by a suture but not occluded. Hemodynamic and cardiac function was monitored using a polygraph. Plasma activity of creatine kinase and the tissue activities of PKA and NOS were measured spectrophotometrically. p-CREB, eNOS and p-eNOS levels were detected using Western blotting. Sizes of the area at risk, the area of no-reflow and the area of necrosis were measured morphologically.

Results: Pretreatment of the animals with simvastatin (SIM, 2 mg/kg, po) before reperfusion significantly decreased the plasma activity of creatine kinase, an index of myocardial necrosis, and reduced the no-reflow size (from 50.4%±2.4% to 36.1%±2.1%, P<0.01) and the infarct size (from 79.0%±2.7% to 64.1%±4.5%, P<0.01). SIM significantly increased the activities of PKA and constitutive NOS, and increased Ser(133) p-CREB protein, Ser(1179) p-eNOS, and Ser(635) p-eNOS in ischemic myocardium. Intravenous infusion of the PKA inhibitor H-89 (1 μg·kg(-1)·min(-1)) partially abrogated the SIM-induced cardioprotection and eNOS phosphorylation. In contrast, intravenous infusion of the eNOS inhibitor L-NNA (10 mg·kg(-1)) completely abrogated the SIM-induced cardioprotection and eNOS phosphorylation during ischemia and reperfusion, but did not affect the activity of PKA.

Conclusion: Pretreatment with a single dose of SIM 2.5 h before reperfusion attenuates myocardial no-reflow and infarction through increasing eNOS phosphorylation at Ser(1179) and Ser(635) that was partially mediated via the PKA signaling pathway.

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References

Birnbaum Y, Lin Y, Ye Y, Merla R, Perez-Polo J, Uretsky B . Pretreatment with high-dose statin, but not low-dose statin, ezetimibe, or the combination of low-dose statin and ezetimibe, limits infarct size in the rat. J Cardiovasc Pharmacol Ther. 2008; 13(1):72-9. DOI: 10.1177/1074248407312839. View

Robinet A, Hoizey G, Millart H . PI 3-kinase, protein kinase C, and protein kinase A are involved in the trigger phase of beta1-adrenergic preconditioning. Cardiovasc Res. 2005; 66(3):530-42. DOI: 10.1016/j.cardiores.2005.02.010. View

Soria L, Gradilone S, Larocca M, Marinelli R . Glucagon induces the gene expression of aquaporin-8 but not that of aquaporin-9 water channels in the rat hepatocyte. Am J Physiol Regul Integr Comp Physiol. 2009; 296(4):R1274-81. DOI: 10.1152/ajpregu.90783.2008. View

Kou R, Michel T . Epinephrine regulation of the endothelial nitric-oxide synthase: roles of RAC1 and beta3-adrenergic receptors in endothelial NO signaling. J Biol Chem. 2007; 282(45):32719-29. DOI: 10.1074/jbc.M706815200. View

Mount P, Kemp B, Power D . Regulation of endothelial and myocardial NO synthesis by multi-site eNOS phosphorylation. J Mol Cell Cardiol. 2006; 42(2):271-9. DOI: 10.1016/j.yjmcc.2006.05.023. View

Michell B, Harris M, Chen Z, Ju H, Venema V, Blackstone M . Identification of regulatory sites of phosphorylation of the bovine endothelial nitric-oxide synthase at serine 617 and serine 635. J Biol Chem. 2002; 277(44):42344-51. DOI: 10.1074/jbc.M205144200. View

Birnbaum Y, Ye Y, Rosanio S, Tavackoli S, Hu Z, Schwarz E . Prostaglandins mediate the cardioprotective effects of atorvastatin against ischemia-reperfusion injury. Cardiovasc Res. 2005; 65(2):345-55. DOI: 10.1016/j.cardiores.2004.10.018. View

Cheng Y, Yang Y, Zhang H, Qian H, Zhao J, Meng X . Pretreatment with Tongxinluo protects porcine myocardium from ischaemia/reperfusion injury through a nitric oxide related mechanism. Chin Med J (Engl). 2009; 122(13):1529-38. View

Ray C, Marshall J . The cellular mechanisms by which adenosine evokes release of nitric oxide from rat aortic endothelium. J Physiol. 2005; 570(Pt 1):85-96. PMC: 1464284. DOI: 10.1113/jphysiol.2005.099390. View

10.

Rikitake Y, Liao J . Rho GTPases, statins, and nitric oxide. Circ Res. 2005; 97(12):1232-5. PMC: 2633589. DOI: 10.1161/01.RES.0000196564.18314.23. View

11.

Makaula S, Lochner A, Genade S, Sack M, Awan M, Opie L . H-89, a non-specific inhibitor of protein kinase A, promotes post-ischemic cardiac contractile recovery and reduces infarct size. J Cardiovasc Pharmacol. 2005; 45(4):341-7. DOI: 10.1097/01.fjc.0000156825.80951.14. View

12.

Resnic F, Wainstein M, Lee M, Behrendt D, Wainstein R, Ohno-Machado L . No-reflow is an independent predictor of death and myocardial infarction after percutaneous coronary intervention. Am Heart J. 2003; 145(1):42-6. DOI: 10.1067/mhj.2003.36. View

13.

Szarszoi O, Maly J, Ostadal P, Netuka I, Besik J, Kolar F . Effect of acute and chronic simvastatin treatment on post-ischemic contractile dysfunction in isolated rat heart. Physiol Res. 2008; 57(5):793-796. DOI: 10.33549/physiolres.931559. View

14.

Zhao J, Yang Y, Cui C, You S, Gao R . Pretreatment with simvastatin reduces myocardial no-reflow by opening mitochondrial K(ATP) channel. Br J Pharmacol. 2006; 149(3):243-9. PMC: 2014278. DOI: 10.1038/sj.bjp.0706862. View

15.

Inserte J, Garcia-Dorado D, Ruiz-Meana M, Agullo L, Pina P, Soler-Soler J . Ischemic preconditioning attenuates calpain-mediated degradation of structural proteins through a protein kinase A-dependent mechanism. Cardiovasc Res. 2004; 64(1):105-14. DOI: 10.1016/j.cardiores.2004.06.001. View

16.

Bellis A, Castaldo D, Trimarco V, Monti M, Chivasso P, Sadoshima J . Cross-talk between PKA and Akt protects endothelial cells from apoptosis in the late ischemic preconditioning. Arterioscler Thromb Vasc Biol. 2009; 29(8):1207-12. DOI: 10.1161/ATVBAHA.109.184135. View

17.

Schulz R, Kelm M, Heusch G . Nitric oxide in myocardial ischemia/reperfusion injury. Cardiovasc Res. 2004; 61(3):402-13. DOI: 10.1016/j.cardiores.2003.09.019. View

18.

Wu Z, Li M, Chen J, Chi Z, Liu J . Involvement of cAMP/cAMP-dependent protein kinase signaling pathway in regulation of Na+,K+-ATPase upon activation of opioid receptors by morphine. Mol Pharmacol. 2005; 69(3):866-76. DOI: 10.1124/mol.105.016501. View

19.

Freireich E, Gehan E, Rall D, Schmidt L, Skipper H . Quantitative comparison of toxicity of anticancer agents in mouse, rat, hamster, dog, monkey, and man. Cancer Chemother Rep. 1966; 50(4):219-44. View

20.

Heusch G, Kleinbongard P, Bose D, Levkau B, Haude M, Schulz R . Coronary microembolization: from bedside to bench and back to bedside. Circulation. 2009; 120(18):1822-36. DOI: 10.1161/CIRCULATIONAHA.109.888784. View