Pharmacological preconditioning and postconditioning with nicorandil attenuates ischemia/reperfusion‑induced myocardial necrosis and apoptosis in hypercholesterolemic rats

Li,Wenna; Wu,Nan; Shu,Wenqi; Jia,Dalin; Jia,Pengyu

doi:10.3892/etm.2015.2782

Pharmacological preconditioning and postconditioning with nicorandil attenuates ischemia/reperfusion‑induced myocardial necrosis and apoptosis in hypercholesterolemic rats

Authors:
- Wenna Li
- Nan Wu
- Wenqi Shu
- Dalin Jia
- Pengyu Jia
View Affiliations

Affiliations: Department of Cardiology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: October 1, 2015 https://doi.org/10.3892/etm.2015.2782
Pages: 2197-2205

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Pharmacological preconditioning and postconditioning may reduce myocardial necrosis and apoptosis during ischemia/reperfusion (I/R), however, hypercholesterolemia interferes with the associated cardioprotective mechanisms. The present study investigated whether pharmacological preconditioning and postconditioning with nicorandil could attenuate myocardial necrosis and apoptosis induced by I/R in hypercholesterolemic rats, and explored the possible mechanisms involved. Male Wistar rats (n=160) were fed normal (normocholesterolemic group, n=10) or high‑cholesterol (hypercholesterolemic group, n=150) diets for 8 weeks. Hearts harvested from the normal and hypercholesterolemic rats were subsequently placed on modified Langendorff perfusion apparatus and 30‑min global ischemia was performed, followed by 120‑min reperfusion. Nicorandil (1, 3, 10, 30, 100 µmol/l), and mitochondrial adenosine triphosphate‑sensitive potassium (mitoKATP) channel blocker 5‑hydroxydecanoic acid sodium salt (5‑HD) (100 µmol/l) or soluble guanylyl cyclase (sGC) blocker 1H‑[1,2,4]oxadiazolo[4,3‑a]quinoxalin‑1‑one (ODQ) (10 µmol/l) were perfused for 10 min, prior to ischemia or at the onset of reperfusion. The myocardial infarct size was determined by triphenyltetrazolium chloride staining, and cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick‑end labeling staining. In order to investigate the potential mechanisms, the expression levels of caspase‑3, B‑cell lymphoma‑2 (Bcl‑2) proteins and Bcl‑2‑associated X protein (Bax) were measured using western blot analysis. The present study demonstrated that, in hypercholesterolemic rats, pharmacological preconditioning and postconditioning with nicorandil decreased I/R‑induced myocardial necrosis and apoptosis in a concentration‑dependent manner. The optimal preconditioning and postconditioning concentration of nicorandil determined to have anti‑infarct and anti‑apoptosis effects was 30 µmol/l, which significantly (P<0.05) reduced the infarct size to 14.88±3.25% and 15.96±3.29%, and attenuated the percentage of cardiomyocyte apoptosis to 25.20±3.93% and 26.18±4.82%, respectively, compared with the I/R group. However, the cardioprotective effects of nicorandil were partially suppressed by cotreatment with 5‑HD or ODQ. Western blot analysis demonstrated that pharmacological preconditioning and postconditioning with nicorandil significantly downregulated caspase‑3 and Bax expression, and upregulated Bcl‑2 expression compared with the I/R group (P<0.05). The results of the present study suggest that pharmacological preconditioning and postconditioning with nicorandil may protect hypercholesterolemic hearts against I/R‑induced necrosis and apoptosis; and the cardioprotective effects of nicorandil may be due to the dual pharmacological mechanisms of opening the mitoKATP channels and a nitric oxide/sGC‑dependent mechanism, and regulation of the expression of caspase‑3, Bax and Bcl‑2.

View Figures

View References

1	Frank A, Bonney M, Bonney S, Weitzel L, Koeppen M and Eckle T: Myocardial ischemia reperfusion injury: From basic science to clinical bedside. Semin Cardiothorac Vasc Anesth. 16:123–132. 2012. View Article : Google Scholar : PubMed/NCBI
2	Hausenloy DJ and Yellon DM: Preconditioning and postconditioning: Underlying mechanisms and clinical application. Atherosclerosis. 204:334–341. 2009. View Article : Google Scholar : PubMed/NCBI
3	Murry CE, Jennings RB and Reimer KA: Preconditioning with ischemia: A delay of lethal cell injury in ischemic myocardium. Circulation. 74:1124–1136. 1986. View Article : Google Scholar : PubMed/NCBI
4	Zhao ZQ, Corvera JS, Haulkos ME, Kerendi F, Wang NP, Guyton RA and Vinten-Joansen J: Inhibition of myocardial injury by ischemic postconditioning during reperfusion comparision with ischemic preconditioning. Am J Physiol Heart Circ Physiol. 285:579–588. 2003. View Article : Google Scholar
5	Gerczuk PZ and Kloner RA: Protecting the heart from ischemia: An update on ischemic and pharmacologic conditioning. Hosp Pract. 39:35–43. 2011. View Article : Google Scholar
6	Andreadou I, Iliodromitis EK, Koufaki M and Kremastinos DT: Pharmacological pre- and post- conditioning agents: Reperfusion-injury of the heart revisited. Mini Rev Med Chem. 8:952–959. 2008. View Article : Google Scholar : PubMed/NCBI
7	Sack MN and Murphy E: The role of comorbidities in cardioprotection. J Cardiovasc Pharmacol Ther. 16:267–272. 2011. View Article : Google Scholar : PubMed/NCBI
8	Balakumar P, Singh H, Singh M and Anand-Srivastava MB: The impairment of preconditioning-mediated cardioprotection in pathological conditions. Pharmacol Res. 60:18–23. 2009. View Article : Google Scholar : PubMed/NCBI
9	Ferdinandy P, Schulz R and Baxter GF: Interaction of cardiovascular risk factors with myocardial ischemia/reperfusion injury, preconditioning, and postconditioning. Pharmacol Rev. 59:418–458. 2007. View Article : Google Scholar : PubMed/NCBI
10	Osipov RM, Bianchi C, Feng J, Clements RT, Liu Y, Robich MP, Glazer HP, Sodha NR and Sellke FW: Effect of hypercholesterolemia on myocardial necrosis and apoptosis in the setting of ischemia-reperfusion. Circulation. 120(Suppl): S22–S30. 2009. View Article : Google Scholar : PubMed/NCBI
11	Wang TD, Chen WJ, Su SS, Lo SC, Lin WW and Lee YT: Increased cardiomyocyte apoptosis following ischemia and reperfusion in diet-induced hypercholesterolemia: Relation to Bcl-2 and Bax proteins and caspase-3 activity. Lipids. 37:385–394. 2002. View Article : Google Scholar : PubMed/NCBI
12	Liu HR, Tao L, Gao E, Qu Y, Lau WB, Lopez BL, Christopher TA, Koch W, Yue TL and Ma XL: Rosiglitazone inhibits hypercholesterolaemia-induced myeloperoxidase upregulation-a novel mechanism for the cardioprotective effects of PPAR agonists. Cardiovasc Res. 81:344–352. 2009. View Article : Google Scholar : PubMed/NCBI
13	Onody A, Csonka C, Giricz Z and Ferdinandy P: Hyperlipidemia induced by a cholesterol-rich diet leads to enhanced peroxynitrite formation in rat hearts. Cardiovasc Res. 58:663–670. 2003. View Article : Google Scholar : PubMed/NCBI
14	Prasan AM, McCarron HC, Zhang Y and Jeremy RW: Myocardial release of nitric oxide during ischaemia and reperfusion: Effects of L-arginine and hypercholesterolaemia. Heart Lung Circ. 16:274–281. 2007. View Article : Google Scholar : PubMed/NCBI
15	Genda S, Miura T, Miki T, Ichikawa Y and Shimamoto K: K(ATP) channel opening is an endogenous mechanism of protection against the no-reflow phenomenon but its function is compromised by hypercholesterolemia. J Am Coll Cardiol. 40:1339–1346. 2002. View Article : Google Scholar : PubMed/NCBI
16	Görbe A, Varga ZV, Kupai K, Bencsik P, Kocsis GF, Csont T, Boengler K, Schulz R and Ferdinandy P: Cholesterol diet leads to attenuation of ischemic preconditioning-induced cardiac protection: the role of connexin 43. Am J Physiol Heart Circ Physiol. 300:H1907–H1913. 2011. View Article : Google Scholar : PubMed/NCBI
17	Wu N, Zhang X, Guan Y, Shu W, Jia P and Jia D: Hypercholesterolemia abrogates the cardioprotection of ischemic postconditioning in isolated rat heart: Roles of glycogen synthase kinase-3β and the mitochondrial permeability transition pore. Cell Biochem Biophys. 69:123–130. 2014. View Article : Google Scholar : PubMed/NCBI
18	Ma LL, Zhang FJ, Qian LB, Kong FJ, Sun JF, Zhou C, Peng YN, Xu HJ, Wang WN, Wen CY, et al: Hypercholesterolemia blocked sevoflurane-induced cardioprotection against ischemia-reperfusion injury by alteration of the MG53/RISK/GSK3β signaling. Int J Cardiol. 168:3671–3678. 2013. View Article : Google Scholar : PubMed/NCBI
19	Horinaka S: Use of nicorandil in cardiovascular disease and its optimization. Drugs. 71:1105–1119. 2011. View Article : Google Scholar : PubMed/NCBI
20	IONA Study Group: Effect of nicorandil on coronary events in patients with stable angina: The Impact Of Nicorandil in Angina (IONA) randomised trial. Lancet. 359:1269–1275. 2002. View Article : Google Scholar : PubMed/NCBI
21	Tsuchida A, Miura T, Tanno M, Sakamoto J, Miki T, Kuno A, Matsumoto T, Ohnuma Y, Ichikawa Y and Shimamoto K: Infarct size limitation by nicorandil: Roles of mitochondrial K(ATP) channels, sarcolemmal K(ATP) channels, and protein kinase C. J Am Coll Cardiol. 40:1523–1530. 2002. View Article : Google Scholar : PubMed/NCBI
22	Akao M, Teshima Y and Marbán E: Antiapoptotic effect of nicorandil mediated by mitochondrial atp-sensitive potassium channels in cultured cardiac myocytes. J Am Coll Cardiol. 40:803–810. 2002. View Article : Google Scholar : PubMed/NCBI
23	Lu C, Minatoguchi S, Arai M, Wang N, Chen XH, Bao N, Kawamura I, Yasuda S, Kobayashi H, Wu DJ, Takemura G and Fujiwara H: Nicorandil improves post-ischemic myocardial dysfunction in association with opening the mitochondrial K(ATP) channels and decreasing hydroxyl radicals in isolated rat hearts. Circ J. 70:1650–1654. 2006. View Article : Google Scholar : PubMed/NCBI
24	Nagata K, Obata K, Odashima M, Yamada A, Somura F, Nishizawa T, Ichihara S, Izawa H, Iwase M, Hayakawa A, Murohara T and Yokota M: Nicorandil inhibits oxidative stress-induced apoptosis in cardiac myocytes through activation of mitochondrial ATP-sensitive potassium channels and a nitrate-like effect. J Mol Cell Cardiol. 35:1505–1512. 2003. View Article : Google Scholar : PubMed/NCBI
25	Nishikawa S, Tatsumi T, Shiraishi J, Matsunaga S, Takeda M, Mano A, Kobara M, Keira N, Okigaki M, Takahashi T and Matsubara H: Nicorandil regulates Bcl-2 family proteins and protects cardiac myocytes against hypoxia-induced apoptosis. J Mol Cell Cardiol. 40:510–519. 2006. View Article : Google Scholar : PubMed/NCBI
26	Wang A, Chen F, Xie Y, Guo Z and Yu Y: Protective mechanism of nicorandil on rat myocardial ischemia-reperfusion. J Cardiovasc Med (Hagerstown). 13:511–515. 2012. View Article : Google Scholar : PubMed/NCBI
27	Sato T, Sasaki N, O'Rourke B and Marbán E: Nicorandil, a potent cardioprotective agent, acts by opening mitochondrial ATP-dependent potassium channels. J Am Coll Cardiol. 35:514–518. 2000. View Article : Google Scholar : PubMed/NCBI
28	Costa AD, Pierre SV, Cohen MV, Downey JM and Garlid KD: cGMP signalling in pre- and post-conditioning: The role of mitochondria. Cardiovasc Res. 77:344–352. 2008. View Article : Google Scholar : PubMed/NCBI
29	Schwemmer M, Sommer O, Koeckerbauer R and Bassenge E: Cardiovascular dysfunction in hypercholesterolemia associated with enhanced formation of AT1-receptor and of eicosanoids. JCardiovasc Pharmacol Ther. 5:59–68. 2000. View Article : Google Scholar
30	Giricz Z, Görbe A, Pipis J, Burley DS, Ferdinandy P and Baxter GF: Hyperlipidaemia induced by a high-cholesterol diet leads to the deterioration of guanosine-3′,5′-cyclic monophosphate/protein kinase G-dependent cardioprotection in rats. Br J Pharmacol. 158:1495–1502. 2009. View Article : Google Scholar : PubMed/NCBI
31	Tang XL, Stein AB, Shirk G and Bolli R: Hypercholesterolemia blunts NO donor-induced late preconditioning against myocardial infarction in conscious rabbits. Basic Res Cardiol. 99:395–403. 2004. View Article : Google Scholar : PubMed/NCBI
32	Sasaki N, Sato T, Ohler A, O'Rourke B and Marbán E: Activation of mitochondrial ATP-dependent potassium channels by nitric oxide. Circulation. 101:439–445. 2000. View Article : Google Scholar : PubMed/NCBI
33	Harada N, Miura T, Dairaku Y, Kametani R, Shibuya M, Wang R, Kawamura S and Matsuzaki M: NO donor-activated PKC-δ plays a pivotal role in ischemic myocardial protection through accelerated opening of mitochondrial K-ATP channels. J Cardiovasc Pharmacol. 44:35–41. 2004. View Article : Google Scholar : PubMed/NCBI
34	Kuno A, Critz SD, Cohen MV and Downey JM: Nicorandil opens mitochondrial K(ATP) channels not only directly but also through a NO-PKG-dependent pathway. Basic Res Cardiol. 102:73–79. 2007. View Article : Google Scholar : PubMed/NCBI
35	Iliodromitis EK, Lazou A and Kremastinos DT: Ischemic preconditioning: Protection against myocardial necrosis and apoptosis. Vasc Health Risk Manag. 3:629–637. 2007.PubMed/NCBI
36	Wang H, Zuo X, Wang Q, Yu Y, Xie L, Wang H, Wu H and Xie W: Nicorandil inhibits hypoxia-induced apoptosis in human pulmonary artery endothelial cells through activation of mitoKATP and regulation of eNOS and the NF-κB pathway. Int J Mol Med. 32:187–194. 2013.PubMed/NCBI
37	Gustafsson AB and Gottlieb RA: Bcl-2 family members and apoptosis, taken to heart. Am J Physiol Cell Physiol. 292:C45–C51. 2007. View Article : Google Scholar : PubMed/NCBI