Abstract
Retinal pigment epithelial (RPE) cells are constantly exposed to oxidative injury while clearing byproducts of photoreceptor turnover, a circumstance thought to be responsible for degenerative retinal diseases. The mechanisms of hydrogen peroxide (H2O2)-induced apoptosis in RPE cells are not fully understood. We studied signal transduction mechanisms of H2O2-induced apoptosis in the human RPE cell line ARPE-19. Activation of two stress kinases (JNK and p38) occurs during H2O2 stimulation, and H2O2-mediated cell death was significantly reduced by their specific inhibition. Exposure to a lethal dose of H2O2 elicited Bax translocation to the mitochondria and release of apoptosis-inducing factor (AIF) from the mitochondria, both of which were abolished by either JNK- or p38-specific inhibitors. Both H2O2-induced cell death and JNK/p38 phosphorylation were partially inhibited by C. difficile toxin B, inhibitor of Rho, Rac, and cdc42. Use of pull-down assays revealed that the small GTPase activated by H2O2 is Rac1. This study is the first to demonstrate that H2O2 induces a Rac1/JNK1/p38 signaling cascade, and that JNK and p38 activation is important for H2O2-induced apoptosis as well as AIF/Bax translocation of RPE cells.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Miceli MV, Liles MR, Newsome DA (1994) Evaluation of oxidative processes in human pigment epithelial cells associated with retinal outer segment phagocytosis. Exp Cell Res 214:242–249
Winkler BS, Boulton ME, Gottsch JD, Sternberg P (1999) Oxidative damage and age-related macular degeneration. Mol Vis 5:32–43
Cai J, Nelson KC, Wu M, Sternberg P Jr, Jones DP (2000) Oxidative damage and protection of the RPE. Prog Retin Eye Res 19:205–221
Voloboueva LA, Liu J, Suh JH, Ames BN, Miller SS (2005) (R)-alpha-lipoic acid protects retinal pigment epithelial cells from oxidative damage. Invest Ophthalmol Vis Sci 46:4302–4310
Kasahara E, Lin LR, Ho YS, Reddy VN (2005) SOD2 protects against oxidation-induced apoptosis in mouse retinal pigment epithelium: implications for age-related macular degeneration. Invest Ophthalmol Vis Sci 46:3426–3434
Strunnikova N, Zhang C, Teichberg D, et al (2004) Survival of retinal pigment epithelium after exposure to prolonged oxidative injury: a detailed gene expression and cellular analysis. Invest Ophthalmol Vis Sci 45:3767–3777
Zhang C, Baffi J, Cousins SW, Csaky KG (2003) Oxidant-induced cell death in retinal pigment epithelium cells mediated through the release of apoptosis-inducing factor. J Cell Sci 116:1915–1923
Jin GF, Hurst JS, Godley BF (2001) Hydrogen peroxide stimulates apoptosis in cultured human retinal pigment epithelial cells. Curr Eye Res 22:165–173
Sparrow JR, Cai B (2001) Blue light-induced apoptosis of A2E-containing RPE: involvement of caspase-3 and protection by Bcl-2. Invest Ophthalmol Vis Sci 42:1356–1362
Howes KA, Liu Y, Dunaief JL, et al (2004) Receptor for advanced glycation end products and age-related macular degeneration. Invest Ophthalmol Vis Sci 45:3713–3720
Del Priore LV, Kuo YH, Tezel TH (2002) Age-related changes in human RPE cell density and apoptosis proportion in situ. Invest Ophthalmol Vis Sci 43:3312–3318
Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME (1995) Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Sci 270:1326–1331
Chang L, Karin M. (2001) Mammalian MAP kinase signalling cascades. Nature 410:37–40
Ono K, Han J (2000) The p38 signal transduction pathway: activation and function. Cell Signal 12:1–13
Davis RJ (2000). Signal transduction by the JNK group of MAP kinases. Cell 103:239–252
Hecquet C, Lefevre G, Valtink M, Engelmann K, Mascarelli F (2003) Activation and role of MAP kinase-dependent pathways in retinal pigment epithelium cells: JNK1, P38 kinase, and cell death. Invest Ophthalmol Vis Sci 44:1320–1329
Susin SA, Lorenzo HK, Zamzami N, et al (1999) Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 397:441–446
Lorenzo HK, Susin SA, Penninger J, Kroemer G (1999) Apoptosis inducing factor (AIF): a phylogenetically old, caspase-independent effector of cell death. Cell Death Differ 6:516–524
Susin SA, Daugas E, Ravagnan L, et al (2000) Two distinct pathways leading to nuclear apoptosis. J Exp Med 192L:571–580
Daugas E, Susin SA, Zamzami N, et al (2000) Mitochondrio-nuclear translocation of AIF in apoptosis and necrosis. FASEB J 14:729–739
Goping IS, Gross A, Lavoie JN, et al (1998) Regulated targeting of BAX to mitochondria. J Cell Bio 143:207–215.
Cregan SP, Fortin A, MacLaurin JG, et al (2002) Apoptosis-inducing factor is involved in the regulation of caspase-independent neuronal cell death. J Cell Biol 158:507–517
Gallo KA, Johnson GL (2002) Mixed-lineage kinase control of JNK and p38 MAPK pathways. Nat Rev Mol Cell Biol 3:663–672
Minden A, Lin A, Claret FX, Abo A, Karin M (1995) Selective activation of the JNK signaling cascade and c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs. Cell 81:1147–1157
Zhang S, Han J, Sells MA, et al (1995) Rho family GTPases regulate p38 mitogen-activated protein kinase through the downstream mediator Pak1. J Biol Chem 270:23934–23936
Bazenet CE, Mota MA, Rubin LL (1998) The small GTP-binding protein Cdc42 is required for nerve growth factor withdrawal-induced neuronal death. Proc Natl Acad Sci USA 95:3984–3989
Friis MB, Friborg CR, Schneider L, et al (2005) Cell shrinkage as a signal to apoptosis in NIH 3T3 fibroblasts. J Physiol 567:427–443
Garg TK, Chang JY (2003) Oxidative stress causes ERK phosphorylation and cell death in cultured retinal pigment epithelium: Prevention of cell death by AG126 and 15-deoxy-delta 12, 14-PGJ2. BMC Ophthalmol 3:5
Tsao YP, Ho TC, Chen SL, Cheng HC (2006) Pigment epithelium-derived factor inhibits oxidative stress-induced cell death by activation of extracellular signal-regulated kinases in cultured retinal pigment epithelial cells. Life Sci (in print).
Prepens U, Just I, von Eichel-Streiber C, Aktories K (1996) Inhibition of Fc epsilon-RI-mediated activation of rat basophilic leukemia cells by Clostridium difficile toxin B (monoglucosyltransferase) J Biol Chem 271:7324–7329
Boisvieux-Ulrich E, Sourdeval M, Marano F (2005) CD437, a synthetic retinoid, induces apoptosis in human respiratory epithelial cells via caspase-independent mitochondrial and caspase-8-dependent pathways both up-regulated by JNK signaling pathway. Exp Cell Res 307:76–90
Stoica BA, Movsesyan VA, Knoblach SM, Faden AI (2005) Ceramide induces neuronal apoptosis through mitogen-activated protein kinases and causes release of multiple mitochondrial proteins. Mol Cell Neurosci 29:355–371
Gross A, Jockel J, Wei MC, Korsmeyer SJ (1998) Enforced dimerization of BAX results in its translocation, mitochondrial dysfunction and apoptosis. EMBO J 17:3878–3885
Bellosillo B, Villamor N, Lopez-Guillermo A, et al (2002) Spontaneous and drug-induced apoptosis is mediated by conformational changes of Bax and Bak in B-cell chronic lymphocytic leukemia. Blood 100:1810–1816
Gross A, Jockel J, Wei MC, Korsmeyer SJ (1998) Enforced dimerization of BAX results in its translocation, mitochondrial dysfunction and apoptosis. EMBO J 17:3878–3885
Lei K, Davis RJ (2003) JNK phosphorylation of Bim-related members of the Bcl2 family induces Bax-dependent apoptosis. Proc Natl Acad Sci USA 100:2432–2437
Geiger RC, Waters CM, Kamp DW, Glucksberg MR (2005) KGF prevents oxygen-mediated damage in ARPE-19 cells. Invest Ophthalmol Vis Sci 46:3435–3442
Bucolo C, Drago F, Lin LR, Reddy VN (2006) Sigma receptor ligands protect human retinal cells against oxidative stress. Neuroreport 17:287–291
Bucolo C, Drago F, Lin LR, Reddy VN (2005) Neuroactive steroids protect retinal pigment epithelium against oxidative stress. Neuroreport 16:1203–1207
Author information
Authors and Affiliations
Corresponding author
Additional information
Y.-C. Yang and T.-C. Ho contributed equally to the work described herein.
Rights and permissions
About this article
Cite this article
Ho, TC., Yang, YC., Cheng, HC. et al. Activation of mitogen-activated protein kinases is essential for hydrogen peroxide -induced apoptosis in retinal pigment epithelial cells. Apoptosis 11, 1899–1908 (2006). https://doi.org/10.1007/s10495-006-9403-6
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10495-006-9403-6