Globular adiponectin protects H9c2 cells from palmitate-induced apoptosis via Akt and ERK1/2 signaling pathways

Lipids in Health and Disease

An Yu Bao²  Wei Wu²  Wen Dai²  Yong Qing Tong²  Kai Sheng Sun²  Hong Yun Zheng²  Yan Li²  Chuan Dong Wei¹ 

[1] Department of Clinical Laboratory, Affiliated Hospital of Youjiang Medical University for nationalities, Baise, Guangxi, 533000, China;Department of Clinical Laboratory, Renmin Hospital of Wuhan University, 99 Ziyang Road, District of Wuchang, Wuhan, Hubei, 430060, China

关键词: ERK1/2;    PI3K/Akt;    Apoptosis;    Palmitate;    H9c2 cells;    Adiponectin;   

Others  :  1160178
DOI  :  10.1186/1476-511X-11-135

 received in 2012-07-26, accepted in 2012-09-29,  发布年份 2012

【 摘 要 】

Background

Cardiomyocytes apoptosis is an important contributor to myocardial dysfunction and heart failure. Adiponectin has cardioprotective effects, potential mechanisms behind it are not clear in cardiomyocytes. The purpose of the study was to investigate whether adiponectin can block palmitate-induced apoptosis and the underlying biochemical mechanism in H9c2 cells.

Methods

H9c2 cells were treated with palmitate presence or absence of 2.5 μg/mL globular adiponectin. The effect on the cell viability of H9c2 cells was evaluated using MTT assay, and cell apoptosis was determined by Hoechst 33342 staining. Protein expression was measured using the western blot method.

Results

Our results showed that the palmitate treatment induced apoptosis in H9c2 cells, which was associated with increasing the level of cleaved caspase-3 and cleaved PARP. Meanwhile, palmitate-induced apoptosis increased the protein level of p-ERK1/2, and decreased the protein level of p-Akt significantly. However, levels of both of these proteins were restored to the normal when pretreated with adiponectin, and followed with the decrease of cleaved caspase-3 and cleaved PARP. In line with these results, the protective effect of adiponectin can be blocked by PI3K/Akt inhibitor LY294002, and palmitate-induced apoptosis can be attenuated by ERK1/2 inhibitor U0126.

Conclusions

Taken together, the present study demonstrated that adiponectin protects H9c2 cells from palmitate-induced apoptosis via PI3K/Akt and ERK1/2 signaling pathways. Our results reveal a link between adiponectin and cardiomyocytes apoptosis, suggesting that adioponectin may be a promising therapeutic for the treatment of lipotoxicity cardiomyopathy.

【 授权许可】

   
2012 Wei et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Zhou YT, Grayburn P, Karim A, Shimabukuro M, Higa M, Baetens D, Orci L, Unger RH: Lipotoxic heart disease in obese rats: implications for human obesity. Proc Natl Acad Sci USA 2000, 97:1784-1789.
• [2]Narula J, Haider N, Virmani R, DiSalvo TG, Kolodgie FD, Hajjar RJ, Schmidt U, Semigran MJ, Dec GW, Khaw BA: Apoptosis in myocytes in end-stage heart failure. N Engl J Med 1996, 335:1182-1189.
• [3]De Vries JE, Vork MM, Roemen TH, De Jong YF, Cleutjens JP, van der Vusse GJ, Van Bilsen M: Saturated but not mono-unsaturated fatty acids induce apoptotic cell death in neonatal rat ventricular myocytes. J Lipid Res 1997, 38:1384-1394.
• [4]Listenberger LL, Ory DS, Schaffer JE: Palmitate-induced apoptosis can occur through a ceramide-independent pathway. J Biol Chem 2001, 276:14890-14895.
• [5]Kong JY, Rabkin SW: Palmitate-induced apoptosis in cardiomyocytes is mediated through alterations in mitochondria: prevention by cyclosporin A. Biochim Biophys Acta 2000, 1485:45-55.
• [6]Baraka A, AbdelGawad H: Targeting apoptosis in the heart of streptozotocin-induced diabetic rats. J Cardiovasc Pharmacol Ther 2010, 15:175-181.
• [7]Scherer PE, Williams S, Fogliano M, Baldini G, Lodish HF: A novel serum protein similar to C1q, produced exclusively in adipocytes. J Biol Chem 1995, 270:26746-26749.
• [8]Maeda K, Okubo K, Shimomura I, Funahashi T, Matsuzawa Y, Matsubara K: cDNA cloning and expression of a novel adipose specific collagen-like factor, apM1 (AdiPose Most abundant Gene transcript 1). Biochem Biophys Res Commun 1996, 221:286-289.
• [9]Fruebis J, Tsao TS, Javorschi S, Ebbets-Reed D, Erickson MR, Yen FT, Bihain BE, Lodish HF: Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice. Proc Natl Acad Sci USA 2001, 98:2005-2010.
• [10]Park M, Youn B, Zheng XL, Wu D, Xu A, Sweeney G: Globular adiponectin, acting via AdipoR1/APPL1, protects H9c2 cells from hypoxia/reoxygenation-induced apoptosis. PLoS One 2011, 6:e19143.
• [11]Pineiro R, Iglesias MJ, Gallego R, Raghay K, Eiras S, Rubio J, Dieguez C, Gualillo O, Gonzalez-Juanatey JR, Lago F: Adiponectin is synthesized and secreted by human and murine cardiomyocytes. FEBS Lett 2005, 579:5163-5169.
• [12]Shibata R, Sato K, Kumada M, Izumiya Y, Sonoda M, Kihara S, Ouchi N, Walsh K: Adiponectin accumulates in myocardial tissue that has been damaged by ischemia-reperfusion injury via leakage from the vascular compartment. Cardiovasc Res 2007, 74:471-479.
• [13]Takahashi T, Yu F, Saegusa S, Sumino H, Nakahashi T, Iwai K, Morimoto S, Kurabayashi M, Kanda T: Impaired expression of cardiac adiponectin in leptin-deficient mice with viral myocarditis. Int Heart J 2006, 47:107-123.
• [14]Tao L, Wang Y, Gao E, Zhang H, Yuan Y, Lau WB, Chan L, Koch WJ, Ma XL: Adiponectin: an indispensable molecule in rosiglitazone cardioprotection following myocardial infarction. Circ Res 2010, 106:409-417.
• [15]Konishi M, Haraguchi G, Ohigashi H, Ishihara T, Saito K, Nakano Y, Isobe M: Adiponectin protects against doxorubicin-induced cardiomyopathy by anti-apoptotic effects through AMPK up-regulation. Cardiovasc Res 2011, 89:309-319.
• [16]Martelli AM, Faenza I, Billi AM, Manzoli L, Evangelisti C, Fala F, Cocco L: Intranuclear 3'-phosphoinositide metabolism and Akt signaling: new mechanisms for tumorigenesis and protection against apoptosis? Cell Signal 2006, 18:1101-1107.
• [17]Lu Z, Xu S: ERK1/2 MAP kinases in cell survival and apoptosis. IUBMB Life 2006, 58:621-631.
• [18]Shibata R, Sato K, Pimentel DR, Takemura Y, Kihara S, Ohashi K, Funahashi T, Ouchi N, Walsh K: Adiponectin protects against myocardial ischemia-reperfusion injury through AMPK- and COX-2-dependent mechanisms. Nat Med 2005, 11:1096-1103.
• [19]Li L, Zhang K, Cai XJ, Feng M, Zhang Y, Zhang M: Adiponectin upregulates prolyl-4-hydroxylase alpha1 expression in interleukin 6-stimulated human aortic smooth muscle cells by regulating ERK 1/2 and Sp1. PLoS One 2011, 6:e22819.
• [20]Cousin SP, Hugl SR, Wrede CE, Kajio H, Myers MG Jr, Rhodes CJ: Free fatty acid-induced inhibition of glucose and insulin-like growth factor I-induced deoxyribonucleic acid synthesis in the pancreatic beta-cell line INS-1. Endocrinology 2001, 142:229-240.
• [21]Nicholson DW, Thornberry NA: Caspases: killer proteases. Trends Biochem Sci 1997, 22:299-306.
• [22]Herceg Z, Wang ZQ: Failure of poly(ADP-ribose) polymerase cleavage by caspases leads to induction of necrosis and enhanced apoptosis. Mol Cell Biol 1999, 19:5124-5133.
• [23]Palanivel R, Fang X, Park M, Eguchi M, Pallan S, De Girolamo S, Liu Y, Wang Y, Xu A, Sweeney G: Globular and full-length forms of adiponectin mediate specific changes in glucose and fatty acid uptake and metabolism in cardiomyocytes. Cardiovasc Res 2007, 75:148-157.
• [24]Baquero AF, Gilbertson TA: Insulin activates epithelial sodium channel (ENaC) via phosphoinositide 3-kinase in mammalian taste receptor cells. Am J Physiol Cell Physiol 2011, 300:C860-C871.
• [25]Diffley JM, Wu M, Sohn M, Song W, Hammad SM, Lyons TJ: Apoptosis induction by oxidized glycated LDL in human retinal capillary pericytes is independent of activation of MAPK signaling pathways. Mol Vis 2009, 15:135-145.
• [26]Garbarino J, Sturley SL: Saturated with fat: new perspectives on lipotoxicity. Curr Opin Clin Nutr Metab Care 2009, 12:110-116.
• [27]Sparagna GC, Hickson-Bick DL, Buja LM, McMillin JB: A metabolic role for mitochondria in palmitate-induced cardiac myocyte apoptosis. Am J Physiol Heart Circ Physiol 2000, 279:H2124-H2132.
• [28]Soldani C, Scovassi AI: Poly(ADP-ribose) polymerase-1 cleavage during apoptosis: an update. Apoptosis 2002, 7:321-328.
• [29]Behrends M, Schulz R, Post H, Alexandrov A, Belosjorow S, Michel MC, Heusch G: Inconsistent relation of MAPK activation to infarct size reduction by ischemic preconditioning in pigs. Am J Physiol Heart Circ Physiol 2000, 279:H1111-H1119.
• [30]Shibata R, Numaguchi Y, Matsushita K, Sone T, Kubota R, Ohashi T, Ishii M, Kihara S, Walsh K, Ouchi N, Murohara T: Usefulness of adiponectin to predict myocardial salvage following successful reperfusion in patients with acute myocardial infarction. Am J Cardiol 2008, 101:1712-1715.
• [31]Pischon T, Girman CJ, Hotamisligil GS, Rifai N, Hu FB, Rimm EB: Plasma adiponectin levels and risk of myocardial infarction in men. JAMA 2004, 291:1730-1737.
• [32]Tao L, Gao E, Jiao X, Yuan Y, Li S, Christopher TA, Lopez BL, Koch W, Chan L, Goldstein BJ, Ma XL: Adiponectin cardioprotection after myocardial ischemia/reperfusion involves the reduction of oxidative/nitrative stress. Circulation 2007, 115:1408-1416.
• [33]Kim JE, Song SE, Kim YW, Kim JY, Park SC, Park YK, Baek SH, Lee IK, Park SY: Adiponectin inhibits palmitate-induced apoptosis through suppression of reactive oxygen species in endothelial cells: involvement of cAMP/protein kinase A and AMP-activated protein kinase. J Endocrinol 2010, 207:35-44.
• [34]Jung TW, Lee YJ, Lee MW, Kim SM: Full-length adiponectin protects hepatocytes from palmitate-induced apoptosis via inhibition of c-Jun NH2 terminal kinase. FEBS J 2009, 276:2278-2284.
• [35]Wijesekara N, Krishnamurthy M, Bhattacharjee A, Suhail A, Sweeney G, Wheeler MB: Adiponectin-induced ERK and Akt phosphorylation protects against pancreatic beta cell apoptosis and increases insulin gene expression and secretion. J Biol Chem 2010, 285:33623-33631.
• [36]Matsui T, Rosenzweig A: Convergent signal transduction pathways controlling cardiomyocyte survival and function: the role of PI 3-kinase and Akt. J Mol Cell Cardiol 2005, 38:63-71.
• [37]Gonon AT, Widegren U, Bulhak A, Salehzadeh F, Persson J, Sjoquist PO, Pernow J: Adiponectin protects against myocardial ischaemia-reperfusion injury via AMP-activated protein kinase, Akt, and nitric oxide. Cardiovasc Res 2008, 78:116-122.
• [38]Maruyama S, Shibata R, Ohashi K, Ohashi T, Daida H, Walsh K, Murohara T, Ouchi N: Adiponectin Ameliorates Doxorubicin-induced Cardiotoxicity through Akt Protein-dependent Mechanism. J Biol Chem 2011, 286:32790-32800.
• [39]Liu J, Mao W, Ding B, Liang CS: ERKs/p53 signal transduction pathway is involved in doxorubicin-induced apoptosis in H9c2 cells and cardiomyocytes. Am J Physiol Heart Circ Physiol 2008, 295:H1956-H1965.
• [40]Wanninger J, Neumeier M, Weigert J, Bauer S, Weiss TS, Schaffler A, Krempl C, Bleyl C, Aslanidis C, Scholmerich J, Buechler C: Adiponectin-stimulated CXCL8 release in primary human hepatocytes is regulated by ERK1/ERK2, p38 MAPK, NF-kappaB, and STAT3 signaling pathways. Am J Physiol Gastrointest Liver Physiol 2009, 297:G611-G618.
• [41]Motobayashi Y, Izawa-Ishizawa Y, Ishizawa K, Orino S, Yamaguchi K, Kawazoe K, Hamano S, Tsuchiya K, Tomita S, Tamaki T: Adiponectin inhibits insulin-like growth factor-1-induced cell migration by the suppression of extracellular signal-regulated kinase 1/2 activation, but not Akt in vascular smooth muscle cells. Hypertens Res 2009, 32:188-193.
• [42]Ren B, Deng Y, Mukhopadhyay A, Lanahan AA, Zhuang ZW, Moodie KL, Mulligan-Kehoe MJ, Byzova TV, Peterson RT, Simons M: ERK1/2-Akt1 crosstalk regulates arteriogenesis in mice and zebrafish. J Clin Invest 2010, 120:1217-1228.