PEDF protects cardiomyocytes by promoting FUNDC1‑mediated mitophagy via PEDF-R under hypoxic condition

Li,Yufeng; Liu,Zhiwei; Zhang,Yiqian; Zhao,Qixiang; Wang,Xiaoyu; Lu,Peng; Zhang,Hao; Wang,Zhu; Dong,Hongyan; Zhang,Zhongming

doi:10.3892/ijmm.2018.3536

PEDF protects cardiomyocytes by promoting FUNDC1‑mediated mitophagy via PEDF-R under hypoxic condition

Authors:
- Yufeng Li
- Zhiwei Liu
- Yiqian Zhang
- Qixiang Zhao
- Xiaoyu Wang
- Peng Lu
- Hao Zhang
- Zhu Wang
- Hongyan Dong
- Zhongming Zhang
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Affiliations: Department of Thoracic Cardiovascular Surgery, Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221006, P.R. China, Research Facility Center for Morphology, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China
Published online on: March 6, 2018 https://doi.org/10.3892/ijmm.2018.3536
Pages: 3394-3404
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Pigment epithelial-derived factor (PEDF) is known to exert diverse physiological activities. Previous studies suggest that hypoxia could induce mitophagy. Astoundingly, under hypoxic condition, we found that PEDF decreased the mitochondrial density of cardiomyocytes. In this study, we evaluated whether PEDF could decrease the mitochondrial density and play a protective role in hypoxic cardiomyocytes via promoting mitophagy. Immunostaining and western blotting were used to analyze mitochondrial density and mitophagy of hypoxic cardiomyocytes. Gas chromatography‑mass spectrometry and ELISA were used to analyze levels of palmitic acid and diacylglycerol. Transmission Electron Microscopy was used to detect mitophagy and the mitochondrial density in adult male Sprague-Dawley rat model of acute myocardial infarction. Compared to the control group, we observed that PEDF decreased mitochondrial density through promoting hypoxic cardiomyocyte mitophagy. PEDF increased the levels of palmitic acid and diacylglycerol, and then upregulated the levels of protein kinase Cα (PKC-α) and its activation. Furthermore, inhibition of PKC-α by Go6976 could effectively suppress PEDF-induced mitophagy. Besides, we found that PEDF promoted FUNDC1-mediated cardiomyocyte mitophagy via ULK1, which depended on the activation of PKC-α. Finally, we discovered that mitophagy was increased and mitochondrial density was reduced in adult male Sprague-Dawley rat model of acute myocardial infarction. We concluded that PEDF promotes mitophagy to protect hypoxic cardiomyocytes, through PEDF/PEDF-R/PA/DAG/PKC-α/ULK1/FUNDC1 pathway.

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1	Lloyd-Jones D, Adams R, Carnethon M, De Simone G, Ferguson TB, Flegal K, Ford E, Furie K, Go A, Greenlund K, et al American Heart Association Statistics Committee and Stroke Statistics Subcommittee: Heart disease and stroke statistics - 2009 update: A report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Circulation. 119:480–486. 2009. View Article : Google Scholar : PubMed/NCBI
2	Begieneman MP, van de Goot FR, Fritz J, Rozendaal R, Krijnen PA and Niessen HW: Validation of ultrastructural analysis of mitochondrial deposits in cardiomyocytes as a method of detecting early acute myocardial infarction in humans. J Forensic Sci. 55:988–992. 2010. View Article : Google Scholar : PubMed/NCBI
3	Wallace DC: A mitochondrial paradigm of metabolic and degenerative diseases, aging, and cancer: A dawn for evolutionary medicine. Annu Rev Genet. 39:359–407. 2005. View Article : Google Scholar : PubMed/NCBI
4	Lin MT and Beal MF: Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature. 443:787–795. 2006. View Article : Google Scholar : PubMed/NCBI
5	Goldman SJ, Taylor R, Zhang Y and Jin S: Autophagy and the degradation of mitochondria. Mitochondrion. 10:309–315. 2010. View Article : Google Scholar : PubMed/NCBI
6	Kawaguchi T, Yamagishi SI and Sata M: Structure-function relationships of PEDF. Curr Mol Med. 10:302–311. 2010. View Article : Google Scholar : PubMed/NCBI
7	Gao X, Zhang H, Zhuang W, Yuan G, Sun T, Jiang X, Zhou Z, Yuan H, Zhang Z and Dong H: PEDF and PEDF-derived peptide 44mer protect cardiomyocytes against hypoxia-induced apoptosis and necroptosis via anti-oxidative effect. Sci Rep. 4:56372014. View Article : Google Scholar : PubMed/NCBI
8	Rodriguez-Enriquez S, Kai Y, Maldonado E, Currin RT and Lemasters JJ: Roles of mitophagy and the mitochondrial permeability transition in remodeling of cultured rat hepatocytes. Autophagy. 5:1099–1106. 2009. View Article : Google Scholar : PubMed/NCBI
9	Jain K, Prasad D, Singh SB and Kohli E: Hypobaric hypoxia imbalances mitochondrial dynamics in rat brain hippocampus. Neurol Res Int. 2015:7420592015.PubMed/NCBI
10	Ikeda Y, Sciarretta S, Nagarajan N, Rubattu S, Volpe M, Frati G and Sadoshima J: New insights into the role of mitochondrial dynamics and autophagy during oxidative stress and aging in the heart. Oxid Med Cell Longev. 2014:2109342014. View Article : Google Scholar : PubMed/NCBI
11	Marzetti E, Csiszar A, Dutta D, Balagopal G, Calvani R and Leeuwenburgh C: Role of mitochondrial dysfunction and altered autophagy in cardiovascular aging and disease: From mechanisms to therapeutics. Am J Physiol Heart Circ Physiol. 305:H459–H476. 2013. View Article : Google Scholar : PubMed/NCBI
12	Dorn GW II: Mitochondrial pruning by Nix and BNip3: An essential function for cardiac-expressed death factors. J Cardiovasc Transl Res. 3:374–383. 2010. View Article : Google Scholar : PubMed/NCBI
13	Hoshino A, Mita Y, Okawa Y, Ariyoshi M, Iwai-Kanai E, Ueyama T, Ikeda K, Ogata T and Matoba S: Cytosolic p53 inhibits Parkin-mediated mitophagy and promotes mitochondrial dysfunction in the mouse heart. Nat Commun. 4:23082013. View Article : Google Scholar : PubMed/NCBI
14	Notari L, Baladron V, Aroca-Aguilar JD, Balko N, Heredia R, Meyer C, Notario PM, Saravanamuthu S, Nueda ML, Sanchez-Sanchez F, et al: Identification of a lipase-linked cell membrane receptor for pigment epithelium-derived factor. J Biol Chem. 281:38022–38037. 2006. View Article : Google Scholar : PubMed/NCBI
15	Zhang H, Sun T, Jiang X, Yu H, Wang M, Wei T, Cui H, Zhuang W, Liu Z, Zhang Z, et al: PEDF and PEDF-derived peptide 44mer stimulate cardiac triglyceride degradation via ATGL. J Transl Med. 13:682015. View Article : Google Scholar : PubMed/NCBI
16	Subramanian P, Locatelli-Hoops S, Kenealey J, DesJardin J, Notari L and Becerra SP: Pigment epithelium-derived factor (PEDF) prevents retinal cell death via PEDF Receptor (PEDF-R): Identification of a functional ligand binding site. J Biol Chem. 288:23928–23942. 2013. View Article : Google Scholar : PubMed/NCBI
17	Zimmermann R, Strauss JG, Haemmerle G, Schoiswohl G, Birner-Gruenberger R, Riederer M, Lass A, Neuberger G, Eisenhaber F, Hermetter A, et al: Fat mobilization in adipose tissue is promoted by adipose triglyceride lipase. Science. 306:1383–1386. 2004. View Article : Google Scholar : PubMed/NCBI
18	Tan SH, Shui G, Zhou J, Li JJ, Bay BH, Wenk MR and Shen HM: Induction of autophagy by palmitic acid via protein kinase C-mediated signaling pathway independent of mTOR (mammalian target of rapamycin). J Biol Chem. 287:14364–14376. 2012. View Article : Google Scholar : PubMed/NCBI
19	Steinberg SF: Structural basis of protein kinase C isoform function. Physiol Rev. 88:1341–1378. 2008. View Article : Google Scholar : PubMed/NCBI
20	Churchill E, Budas G, Vallentin A, Koyanagi T and Mochly-Rosen D: PKC isozymes in chronic cardiac disease: Possible therapeutic targets? Annu Rev Pharmacol Toxicol. 48:569–599. 2008. View Article : Google Scholar
21	Ping P, Zhang J, Qiu Y, Tang XL, Manchikalapudi S, Cao X and Bolli R: Ischemic preconditioning induces selective translocation of protein kinase C isoforms epsilon and eta in the heart of conscious rabbits without subcellular redistribution of total protein kinase C activity. Circ Res. 81:404–414. 1997. View Article : Google Scholar : PubMed/NCBI
22	Hambleton M, Hahn H, Pleger ST, Kuhn MC, Klevitsky R, Carr AN, Kimball TF, Hewett TE, Dorn GW II, Koch WJ, et al: Pharmacological- and gene therapy-based inhibition of protein kinase Calpha/beta enhances cardiac contractility and attenuates heart failure. Circulation. 114:574–582. 2006. View Article : Google Scholar : PubMed/NCBI
23	Narendra D, Tanaka A, Suen DF and Youle RJ: Parkin is recruited selectively to impaired mitochondria and promotes their autophagy. J Cell Biol. 183:795–803. 2008. View Article : Google Scholar : PubMed/NCBI
24	Liu L, Feng D, Chen G, Chen M, Zheng Q, Song P, Ma Q, Zhu C, Wang R, Qi W, et al: Mitochondrial outer-membrane protein FUNDC1 mediates hypoxia-induced mitophagy in mammalian cells. Nat Cell Biol. 14:177–185. 2012. View Article : Google Scholar : PubMed/NCBI
25	Novak I, Kirkin V, McEwan DG, Zhang J, Wild P, Rozenknop A, Rogov V, Löhr F, Popovic D, Occhipinti A, et al: Nix is a selective autophagy receptor for mitochondrial clearance. EMBO Rep. 11:45–51. 2010. View Article : Google Scholar :
26	Wu W, Tian W, Hu Z, Chen G, Huang L, Li W, Zhang X, Xue P, Zhou C, Liu L, et al: ULK1 translocates to mitochondria and phosphorylates FUNDC1 to regulate mitophagy. EMBO Rep. 15:566–575. 2014. View Article : Google Scholar : PubMed/NCBI
27	Zhang H, Wang Z, Feng SJ, Xu L, Shi HX, Chen LL, Yuan GD, Yan W, Zhuang W, Zhang YQ, et al: PEDF improves cardiac function in rats with acute myocardial infarction via inhibiting vascular permeability and cardiomyocyte apoptosis. Int J Mol Sci. 16:5618–5634. 2015. View Article : Google Scholar : PubMed/NCBI
28	Khan M, Meduru S, Gogna R, Madan E, Citro L, Kuppusamy ML, Sayyid M, Mostafa M, Hamlin RL and Kuppusamy P: Oxygen cycling in conjunction with stem cell transplantation induces NOS3 expression leading to attenuation of fibrosis and improved cardiac function. Cardiovasc Res. 93:89–99. 2012. View Article : Google Scholar
29	Fukuhara S, Tomita S, Yamashiro S, Morisaki T, Yutani C, Kitamura S and Nakatani T: Direct cell-cell interaction of cardiomyocytes is key for bone marrow stromal cells to go into cardiac lineage in vitro. J Thorac Cardiovasc Surg. 125:1470–1480. 2003. View Article : Google Scholar : PubMed/NCBI
30	Luedde M, Lutz M, Carter N, Sosna J, Jacoby C, Vucur M, Gautheron J, Roderburg C, Borg N, Reisinger F, et al: RIP3, a kinase promoting necroptotic cell death, mediates adverse remodelling after myocardial infarction. Cardiovasc Res. 103:206–216. 2014. View Article : Google Scholar : PubMed/NCBI
31	Fujimura M, Morita-Fujimura Y, Kawase M, Copin JC, Calagui B, Epstein CJ and Chan PH: Manganese superoxide dismutase mediates the early release of mitochondrial cytochrome c and subsequent DNA fragmentation after permanent focal cerebral ischemia in mice. J Neurosci. 19:3414–3422. 1999.PubMed/NCBI
32	Akbari M, Otterlei M, Peña-Diaz J and Krokan HE: Different organization of base excision repair of uracil in DNA in nuclei and mitochondria and selective upregulation of mitochondrial uracil-DNA glycosylase after oxidative stress. Neuroscience. 145:1201–1212. 2007. View Article : Google Scholar
33	Wang X, Zhang Y, Lu P, Zhang H, Li Y, Dong H and Zhang Z: PEDF attenuates hypoxia-induced apoptosis and necrosis in H9c2 cells by inhibiting p53 mitochondrial translocation via PEDF-R. Biochem Biophys Res Commun. 465:394–401. 2015. View Article : Google Scholar : PubMed/NCBI
34	Murray AJ: Metabolic adaptation of skeletal muscle to high altitude hypoxia: How new technologies could resolve the controversies. Genome Med. 1:1172009. View Article : Google Scholar
35	Karbowski M and Youle RJ: Dynamics of mitochondrial morphology in healthy cells and during apoptosis. Cell Death Differ. 10:870–880. 2003. View Article : Google Scholar : PubMed/NCBI
36	Li W, Zhang X, Zhuang H, Chen HG, Chen Y, Tian W, Wu W, Li Y, Wang S, Zhang L, et al: MicroRNA-137 is a novel hypoxia-responsive microRNA that inhibits mitophagy via regulation of two mitophagy receptors FUNDC1 and NIX. J Biol Chem. 289:10691–10701. 2014. View Article : Google Scholar : PubMed/NCBI
37	Listenberger LL, Han X, Lewis SE, Cases S, Farese RV Jr, Ory DS and Schaffer JE: Triglyceride accumulation protects against fatty acid-induced lipotoxicity. Proc Natl Acad Sci USA. 100:3077–3082. 2003. View Article : Google Scholar : PubMed/NCBI
38	Coleman RA and Mashek DG: Mammalian triacylglycerol metabolism: Synthesis, lipolysis, and signaling. Chem Rev. 111:6359–6386. 2011. View Article : Google Scholar : PubMed/NCBI
39	Li LO, Klett EL and Coleman RA: Acyl-CoA synthesis, lipid metabolism and lipotoxicity. Biochim Biophys Acta. 1801:246–251. 2010. View Article : Google Scholar :
40	Newton AC: Protein kinase C: Structural and spatial regulation by phosphorylation, cofactors, and macromolecular interactions. Chem Rev. 101:2353–2364. 2001. View Article : Google Scholar : PubMed/NCBI
41	Ferretti G: Limiting factors to oxygen transport on Mount Everest 30 years after: A critique of Paolo Cerretelli's contribution to the study of altitude physiology. Eur J Appl Physiol. 90:344–350. 2003. View Article : Google Scholar : PubMed/NCBI
42	Levett DZ, Radford EJ, Menassa DA, Graber EF, Morash AJ, Hoppeler H, Clarke K, Martin DS, Ferguson-Smith AC, Montgomery HE, et al: Caudwell Xtreme Everest Research Group: Acclimatization of skeletal muscle mitochondria to high-altitude hypoxia during an ascent of Everest. FASEB J. 26:1431–1441. 2012. View Article : Google Scholar
43	Howald H and Hoppeler H: Performing at extreme altitude: Muscle cellular and subcellular adaptations. Eur J Appl Physiol. 90:360–364. 2003. View Article : Google Scholar : PubMed/NCBI
44	Kim I, Rodriguez-Enriquez S and Lemasters JJ: Selective degradation of mitochondria by mitophagy. Arch Biochem Biophys. 462:245–253. 2007. View Article : Google Scholar : PubMed/NCBI
45	Ashrafi G and Schwarz TL: The pathways of mitophagy for quality control and clearance of mitochondria. Cell Death Differ. 20:31–42. 2013. View Article : Google Scholar
46	Mammucari C and Rizzuto R: Signaling pathways in mitochondrial dysfunction and aging. Mech Ageing Dev. 131:536–543. 2010. View Article : Google Scholar : PubMed/NCBI
47	Piro S, Anello M, Di Pietro C, Lizzio MN, Patanè G, Rabuazzo AM, Vigneri R, Purrello M and Purrello F: Chronic exposure to free fatty acids or high glucose induces apoptosis in rat pancreatic islets: Possible role of oxidative stress. Metabolism. 51:1340–1347. 2002. View Article : Google Scholar : PubMed/NCBI
48	Narendra DP, Jin SM, Tanaka A, Suen DF, Gautier CA, Shen J, Cookson MR and Youle RJ: PINK1 is selectively stabilized on impaired mitochondria to activate Parkin. PLoS Biol. 8:e10002982010. View Article : Google Scholar : PubMed/NCBI
49	Sandoval H, Thiagarajan P, Dasgupta SK, Schumacher A, Prchal JT, Chen M and Wang J: Essential role for Nix in autophagic maturation of erythroid cells. Nature. 454:232–235. 2008. View Article : Google Scholar : PubMed/NCBI