Protective effects of SS31 on t-BHP induced oxidative damage in 661W cells

The present study aimed to investigate the ability of SS31, a novel mitochondria-targeted peptide to protect against t-BHP-induced mitochondrial dysfunction and apoptosis in 661W cell lines. The 661W cells were treated with various concentrations of SS-31 and an MTT assay was used to determine cell viability. The expression of nitrotyrosine and 8-hydroxydeoxyguanosine (8-OHdG) was detected using immunofluorescent staining. Apoptosis were assessed using Hoechst staining and an annexin V/propidium iodide flow cytometer. Reactive oxygen species (ROS) were detected using MitoSOX™ with confocal microscopy. Changes in mitochondrial membrane potential were analyzed using flow cytometry. In addition, the release of cytochrome c was analyzed using confocal microscopy. The viability of the cells improved following treatment with SS31 between 100 nM and 1 µM, compared with untreated control group. Compared with the t-BHP treatment group (20.0±3.8%), the number of annexin V-positive cells decreased dose-dependently to 13.6±2.6, 9.8±0.5 and 7.4±2.0% in the SS-31 treated group at concentrations of 10 nM, 100 nM and 1 µM, respectively. Treatment with SS-31 significantly prevented the t-BHP-induced expression of nitrotyrosine and 8-OHdG, decreased the quantity of mitochondrial ROS, increased mitochondrial potential, and prevented the release of cytochrome c from mitochondria into the cytoplasm. Therefore, the SS31 mitochondria-targeted peptide protected the 661W cells from the sustained oxidative stress induced by t-BHP.

[1]  鍾景光.  Molecular Medicine Reports , 2015 .

[2]  M. A. Akbarsha,et al.  Verrucarin A Induces Apoptosis Through ROS‐Mediated EGFR/MAPK/Akt Signaling Pathways in MDA‐MB‐231 Breast Cancer Cells , 2014, Journal of cellular biochemistry.

[3]  M. Sikorska,et al.  Nanomicellar formulation of coenzyme Q10 (Ubisol-Q10) effectively blocks ongoing neurodegeneration in the mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine model: potential use as an adjuvant treatment in Parkinson's disease , 2014, Neurobiology of Aging.

[4]  X. Chen,et al.  Mitochondria-targeted antioxidant peptide SS31 protects the retinas of diabetic rats. , 2013, Current molecular medicine.

[5]  P. Reddy,et al.  Impaired mitochondrial biogenesis, defective axonal transport of mitochondria, abnormal mitochondrial dynamics and synaptic degeneration in a mouse model of Alzheimer's disease. , 2011, Human molecular genetics.

[6]  J. Ge,et al.  Mitochondria-targeted peptide MTP-131 alleviates mitochondrial dysfunction and oxidative damage in human trabecular meshwork cells. , 2011, Investigative ophthalmology & visual science.

[7]  B. Chazotte.  Labeling mitochondria with JC-1. , 2011, Cold Spring Harbor protocols.

[8]  Xiaoling Liang,et al.  Mitochondria-targeted antioxidant peptide SS31 attenuates high glucose-induced injury on human retinal endothelial cells. , 2011, Biochemical and biophysical research communications.

[9]  R. Martins,et al.  Lipoic acid as an anti-inflammatory and neuroprotective treatment for Alzheimer's disease. , 2008, Advanced drug delivery reviews.

[10]  K. Groebe,et al.  Nonenzymatic posttranslational protein modifications in ageing , 2008, Experimental Gerontology.

[11]  T. Veen,et al.  Significant photoreceptor rescue by treatment with a combination of antioxidants in an animal model for retinal degeneration , 2007, Neuroscience.

[12]  H. Szeto,et al.  A Novel Cell-permeable Antioxidant Peptide, SS31, Attenuates Ischemic Brain Injury by Down-regulating CD36* , 2007, Journal of Biological Chemistry.

[13]  H. Szeto.  Mitochondria-targeted peptide antioxidants: Novel neuroprotective agents , 2006, The AAPS Journal.

[14]  T. Cotter,et al.  Ceramide is the key mediator of oxidative stress‐induced apoptosis in retinal photoreceptor cells , 2006, Journal of neurochemistry.

[15]  M. Beal,et al.  Cell‐permeable peptide antioxidants as a novel therapeutic approach in a mouse model of amyotrophic lateral sclerosis , 2006 .

[16]  P. Campochiaro,et al.  Antioxidants reduce cone cell death in a model of retinitis pigmentosa. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[17]  L. H. van den Berg,et al.  Intake of polyunsaturated fatty acids and vitamin E reduces the risk of developing amyotrophic lateral sclerosis , 2006, Journal of Neurology, Neurosurgery & Psychiatry.

[18]  H. Szeto,et al.  Mitochondria-targeted peptide prevents mitochondrial depolarization and apoptosis induced by tert-butyl hydroperoxide in neuronal cell lines. , 2005, Biochemical pharmacology.

[19]  P. Campochiaro,et al.  Oxidative damage is a potential cause of cone cell death in retinitis pigmentosa , 2005, Journal of cellular physiology.

[20]  Roberto Pastor-Barriuso,et al.  Meta-Analysis: High-Dosage Vitamin E Supplementation May Increase All-Cause Mortality , 2005, Annals of Internal Medicine.

[21]  Chris A. Johnson,et al.  Further evaluation of docosahexaenoic acid in patients with retinitis pigmentosa receiving vitamin A treatment: subgroup analyses. , 2004, Archives of ophthalmology.

[22]  H. Szeto,et al.  Cell-permeable Peptide Antioxidants Targeted to Inner Mitochondrial Membrane inhibit Mitochondrial Swelling, Oxidative Cell Death, and Reperfusion Injury* , 2004, Journal of Biological Chemistry.

[23]  M. Naash,et al.  Expression of cone-photoreceptor-specific antigens in a cell line derived from retinal tumors in transgenic mice. , 2004, Investigative ophthalmology & visual science.

[24]  Robin A. J. Smith,et al.  Mitochondria‐targeted antioxidants protect Friedreich Ataxia fibroblasts from endogenous oxidative stress more effectively than untargeted antioxidants , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  N. Rotstein,et al.  Protective effect of docosahexaenoic acid on oxidative stress-induced apoptosis of retina photoreceptors. , 2003, Investigative ophthalmology & visual science.

[26]  J. Paul Robinson,et al.  Mitochondrial Complex I Inhibitor Rotenone Induces Apoptosis through Enhancing Mitochondrial Reactive Oxygen Species Production* , 2003, The Journal of Biological Chemistry.

[27]  H. Szeto,et al.  A highly potent peptide analgesic that protects against ischemia-reperfusion-induced myocardial stunning. , 2002, American journal of physiology. Heart and circulatory physiology.

[28]  C. Craft,et al.  Retinoic acid upregulates cone arrestin expression in retinoblastoma cells through a Cis element in the distal promoter region. , 2002, Investigative ophthalmology & visual science.

[29]  B. Aggarwal,et al.  Bcl-2 overexpression protects photooxidative stress-induced apoptosis of photoreceptor cells via NF-kappaB preservation. , 2001, Biochemical and biophysical research communications.

[30]  M. Benito,et al.  Reactive oxygen species (ROS) mediates the mitochondrial‐dependent apoptosis induced by transforming growth factor ß in fetal hepatocytes , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[31]  Robin A. J. Smith,et al.  Selective Targeting of a Redox-active Ubiquinone to Mitochondria within Cells , 2001, The Journal of Biological Chemistry.

[32]  F. Levi-Schaffer,et al.  Role of reactive oxygen species (ROS) in apoptosis induction , 2000, Apoptosis.

[33]  Dao-Yi Yu,et al.  Intraretinal oxygen levels before and after photoreceptor loss in the RCS rat. , 2000, Investigative ophthalmology & visual science.

[34]  J. Stone,et al.  Tissue oxygen during a critical developmental period controls the death and survival of photoreceptors. , 1997, Investigative ophthalmology & visual science.

[35]  B. Herman,et al.  Mitochondrial permeability transition in hepatocytes induced by t-BuOOH: NAD(P)H and reactive oxygen species. , 1997, The American journal of physiology.

[36]  T K Goldstick,et al.  Oxygen consumption in the inner and outer retina of the cat. , 1995, Investigative ophthalmology & visual science.

[37]  J. Schulz,et al.  Inhibition of Neuronal Nitric Oxide Synthase by 7‐Nitroindazole Protects Against MPTP‐Induced Neurotoxicity in Mice , 1995, Journal of neurochemistry.

[38]  J. Nathans,et al.  Apoptotic photoreceptor cell death in mouse models of retinitis pigmentosa. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[39]  B. W. Nicholson,et al.  A randomized trial of vitamin A and vitamin E supplementation for retinitis pigmentosa. , 1993, Archives of ophthalmology.

[40]  Y. Hao,et al.  Apoptosis: Final common pathway of photoreceptor death in rd, rds, and mutant mice , 1993, Neuron.

[41]  M. Al-Ubaidi,et al.  Bilateral retinal and brain tumors in transgenic mice expressing simian virus 40 large T antigen under control of the human interphotoreceptor retinoid-binding protein promoter , 1992, The Journal of cell biology.

[42]  G. Chader,et al.  Effects of butyrate, retinol, and retinoic acid on human Y-79 retinoblastoma cells growing in monolayer cultures. , 1984, Journal of the National Cancer Institute.

[43]  M. Beal,et al.  Cell-permeable peptide antioxidants as a novel therapeutic approach in a mouse model of amyotrophic lateral sclerosis. , 2006, Journal of neurochemistry.

[44]  M. Beal,et al.  Coenzyme Q10 in the central nervous system and its potential usefulness in the treatment of neurodegenerative diseases. , 1997, Molecular aspects of medicine.

[45]  M. Bottone,et al.  Detection of apoptotic cells by annexin V labeling at electron microscopy. , 1997, European journal of histochemistry : EJH.

[46]  Y. Hao,et al.  Apoptosis: final common pathway of photoreceptor death in rd, rds, and rhodopsin mutant mice. , 1993, Neuron.

[47]  D D Clowes,et al.  A randomized trial of vitamin A and vitamin E supplementation for retinitis pigmentosa. , 1993, Archives of ophthalmology.

[48]  S. Liebowitz.  Retinitis pigmentosa. , 1979, Journal - American Intra-Ocular Implant Society.