Resveratrol attenuates hypoxia/reoxygenation‑induced Ca2+ overload by inhibiting the Wnt5a/Frizzled‑2 pathway in rat H9c2 cells

Wu,Xiang; Zhou,Shanshan; Zhu,Ning; Wang,Xianbao; Jin,Wen; Song,Xudong; Chen,Aihua

doi:10.3892/mmr.2014.2488

Resveratrol attenuates hypoxia/reoxygenation‑induced Ca2+ overload by inhibiting the Wnt5a/Frizzled‑2 pathway in rat H9c2 cells

Authors:
- Xiang Wu
- Shanshan Zhou
- Ning Zhu
- Xianbao Wang
- Wen Jin
- Xudong Song
- Aihua Chen
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Affiliations: Department of Cardiology, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510282, P.R. China, Department of Pharmacy, The Third People's Hospital of Nanhai District, Foshan, Guangdong 528244, P.R. China, Department of Cardiology, Guangdong No. 2 Provincial People's Hospital of Southern Medical University, Guangzhou, Guangdong 510317, P.R. China
Published online on: August 14, 2014 https://doi.org/10.3892/mmr.2014.2488
Pages: 2542-2548

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Abstract

Resveratrol is able to protect myocardial cells from ischemia/reperfusion‑induced injury. However, the mechanism has yet to be fully elucidated. In the present study, it is reported that resveratrol has a critical role in the control of Ca2+ overload, which is the primary underlying cause of ischemia/reperfusion injury. Hypoxia/reoxygenation (H/R) treatment decreased the cell viability and increased the apoptosis of H9c2 cells, whereas the caspase‑3 and intracellular Ca2+ levels were greatly elevated compared with the control group. Treatment of H9c2 cells with resveratrol (5, 15 and 30 µM) reduced caspase‑3 expression and cardiomyocyte apoptosis in a dose‑dependent manner, and the intracellular Ca2+ overload was also significantly decreased. Furthermore, Frizzled‑2 and Wnt5a belong to the non‑canonical Wnt/Ca2+ pathway, which have been demonstrated to be responsible for Ca2+ overload, and were thus detected in the present study. The results indicated that both the mRNA and protein expression levels of Frizzled‑2 and Wnt5a in H/R‑induced H9c2 cells were markedly increased compared with the levels found in normal cells, and treatment with resveratrol (5, 15 and 30 µM) significantly reduced the expression of Frizzled‑2 and Wnt5a compared with the H/R group. The results indicated that resveratrol protected myocardial cells from H/R injury by inhibiting the Ca2+ overload through suppression of the Wnt5a/Frizzled‑2 pathway.

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1	Boateng S and Sanborn T: Acute myocardial infarction. Dis Mon. 59:83–96. 2013. View Article : Google Scholar
2	Yellon DM and Hausenloy DJ: Myocardial reperfusion injury. N Engl J Med. 357:1121–1135. 2007. View Article : Google Scholar : PubMed/NCBI
3	Turer AT and Hill JA: Pathogenesis of myocardial ischemia-reperfusion injury and rationale for therapy. Am J Cardiol. 106:360–368. 2010. View Article : Google Scholar : PubMed/NCBI
4	Weiss JN, Korge P, Honda HM and Ping P: Role of the mitochondrial permeability transition in myocardial disease. Circ Res. 93:292–301. 2003. View Article : Google Scholar : PubMed/NCBI
5	Fulda S and Debatin KM: Sensitization for tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by the chemopreventive agent resveratrol. Cancer Res. 64:337–346. 2004. View Article : Google Scholar
6	Genade T and Lang DM: Resveratrol extends lifespan and preserves glia but not neurons of the Nothobranchius guentheri optic tectum. Exp Gerontol. 48:202–212. 2013. View Article : Google Scholar : PubMed/NCBI
7	Wang SJ, Bo QY, Zhao XH, Yang X, Chi ZF and Liu XW: Resveratrol pre-treatment reduces early inflammatory responses induced by status epilepticus via mTOR signaling. Brain Res. 1492:122–129. 2013. View Article : Google Scholar : PubMed/NCBI
8	Hao HD and He LR: Mechanisms of cardiovascular protection by resveratrol. J Med Food. 7:290–298. 2004. View Article : Google Scholar : PubMed/NCBI
9	Wu JM and Hsieh TC: Resveratrol: a cardioprotective substance. Ann N Y Acad Sci. 1215:16–21. 2011. View Article : Google Scholar : PubMed/NCBI
10	Ungvari Z, Labinskyy N, Mukhopadhyay P, et al: Resveratrol attenuates mitochondrial oxidative stress in coronary arterial endothelial cells. Am J Physiol Heart Circ Physiol. 297:H1876–H1881. 2009. View Article : Google Scholar : PubMed/NCBI
11	Shen M, Wu RX, Zhao L, et al: Resveratrol attenuates ischemia/reperfusion injury in neonatal cardiomyocytes and its underlying mechanism. PLoS One. 7:e512232012. View Article : Google Scholar : PubMed/NCBI
12	Kuhl M, Sheldahl LC, Park M, Miller JR and Moon RT: The Wnt/Ca²⁺ pathway: a new vertebrate Wnt signaling pathway takes shape. Trends Genet. 16:279–283. 2000. View Article : Google Scholar
13	Slusarski DC, Corces VG and Moon RT: Interaction of Wnt and a Frizzled homologue triggers G-protein-linked phosphatidylinositol signalling. Nature. 390:410–413. 1997. View Article : Google Scholar : PubMed/NCBI
14	Sheldahl LC, Park M, Malbon CC and Moon RT: Protein kinase C is differentially stimulated by Wnt and Frizzled homologs in a G-protein-dependent manner. Curr Biol. 9:695–698. 1999. View Article : Google Scholar : PubMed/NCBI
15	Zhou SS, He F, Chen AH, Hao PY and Song XD: Suppression of rat Frizzled-2 attenuates hypoxia/reoxygenation-induced Ca²⁺ accumulation in rat H9c2 cells. Exp Cell Res. 318:1480–1491. 2012. View Article : Google Scholar : PubMed/NCBI
16	Chen M, Sun HY, Li SJ, Das M, Kong JM and Gao TM: Nitric oxide as an upstream signal of p38 mediates hypoxia/reoxygenation-induced neuronal death. Neurosignals. 17:162–168. 2009. View Article : Google Scholar : PubMed/NCBI
17	Shin EJ, Schram K, Zheng XL and Sweeney G: Leptin attenuates hypoxia/reoxygenation-induced activation of the intrinsic pathway of apoptosis in rat H9c2 cells. J Cell Physiol. 221:490–497. 2009. View Article : Google Scholar : PubMed/NCBI
18	Zhang C, Lin G, Wan W, et al: Resveratrol, a polyphenol phytoalexin, protects cardiomyocytes against anoxia/reoxygenation injury via the TLR4/NF-κB signaling pathway. Int J Mol Med. 29:557–563. 2012.PubMed/NCBI
19	Porter AG and Jänicke RU: Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 6:99–104. 1999. View Article : Google Scholar : PubMed/NCBI
20	Kajstura J, Cheng W, Reiss K, et al: Apoptotic and necrotic myocyte cell deaths are independent contributing variables of infarct size in rats. Lab Invest. 74:86–107. 1996.PubMed/NCBI
21	Robich MP, Osipov RM, Nezafat R, et al: Resveratrol improves myocardial perfusion in a swine model of hypercholesterolemia and chronic myocardial ischemia. Circulation. 122(Suppl): S142–S149. 2010. View Article : Google Scholar : PubMed/NCBI
22	Burstein B, Maguy A, Clément R, et al: Effects of resveratrol (trans-3,5,4′-trihydroxystilbene) treatment on cardiac remodeling following myocardial infarction. J Pharmacol Exp Ther. 323:916–923. 2007.
23	Eguchi M, Liu Y, Shin EJ and Sweeney G: Leptin protects H9c2 rat cardiomyocytes from H2O2-induced apoptosis. FEBS J. 275:3136–3144. 2008. View Article : Google Scholar : PubMed/NCBI
24	Das S, Alagappan VK, Bagchi D, et al: Coordinated induction of iNOS-VEGF-KDR-eNOS after resveratrol consumption: a potential mechanism for resveratrol preconditioning of the heart. Vascul Pharmacol. 42:281–289. 2005. View Article : Google Scholar
25	Das S, Tosaki A, Bagchi D, et al: Resveratrol-mediated activation of cAMP response element-binding protein through adenosine A3 receptor by Akt-dependent and -independent pathways. J Pharmacol Exp Ther. 314:762–769. 2005. View Article : Google Scholar
26	Moens AL, Claeys MJ, Timmermans JP and Vrints CJ: Myocardial ischemia/reperfusion-injury, a clinical view on a complex pathophysiological process. Int J Cardiol. 100:179–190. 2005. View Article : Google Scholar : PubMed/NCBI
27	Bradamante S, Barenghi L, Piccinini F, et al: Resveratrol provides late-phase cardioprotection by means of a nitric oxide-and adenosine-mediated mechanism. Eur J Pharmacol. 465:115–123. 2003. View Article : Google Scholar
28	Das S, Cordis GA, Maulik N and Das DK: Pharmacological preconditioning with resveratrol: role of CREB-dependent Bcl-2 signaling via adenosine A3 receptor activation. Am J Physiol Heart Circ Physiol. 288:H328–H335. 2005. View Article : Google Scholar : PubMed/NCBI
29	Shimamoto A, Chong AJ, Yada M, et al: Inhibition of Toll-like receptor 4 with eritoran attenuates myocardial ischemia-reperfusion injury. Circulation. 114(Suppl): I270–I274. 2006. View Article : Google Scholar : PubMed/NCBI
30	Dudley J, Das S, Mukherjee S and Das DK: Resveratrol, a unique phytoalexin present in red wine, delivers either survival signal or death signal to the ischemic myocardium depending on dose. J Nutr Biochem. 20:443–452. 2009. View Article : Google Scholar
31	Gasser RN: The interdependence of hypertension, calcium overload, and coronary spasm in the development of myocardial infarction. Angiology. 39:761–772. 1988. View Article : Google Scholar : PubMed/NCBI
32	Borutaite V and Brown GC: Mitochondria in apoptosis of ischemic heart. FEBS Lett. 541:1–5. 2003. View Article : Google Scholar : PubMed/NCBI
33	Hausenloy DJ and Yellon DM: The mitochondrial permeability transition pore: its fundamental role in mediating cell death during ischaemia and reperfusion. J Mol Cell Cardiol. 35:339–341. 2003. View Article : Google Scholar : PubMed/NCBI
34	Borutaite V, Jekabsone A, Morkuniene R and Brown GC: Inhibition of mitochondrial permeability transition prevents mitochondrial dysfunction, cytochrome c release and apoptosis induced by heart ischemia. J Mol Cell Cardiol. 35:357–366. 2003. View Article : Google Scholar
35	Pei JM, Kravtsov GM, Wu S, et al: Calcium homeostasis in rat cardiomyocytes during chronic hypoxia: a time course study. Am J Physiol Cell Physiol. 285:C1420–C1428. 2003. View Article : Google Scholar : PubMed/NCBI
36	Penna C, Perrelli MG and Pagliaro P: Mitochondrial pathways, permeability transition pore, and redox signaling in cardioprotection: therapeutic implications. Antioxid Redox Signal. 18:556–599. 2013. View Article : Google Scholar : PubMed/NCBI
37	Koval A, Purvanov V, Egger-Adam D and Katanaev VL: Yellow submarine of the Wnt/Frizzled signaling: submerging from the G protein harbor to the targets. Biochem Pharmacol. 82:1311–1319. 2011. View Article : Google Scholar : PubMed/NCBI
38	Clevers H and Nusse R: Wnt/β-catenin signaling and disease. Cell. 149:1192–1205. 2012.