Neuroprotection of a Novel Cyclopeptide C*HSDGIC* from the Cyclization of PACAP (1–5) in Cellular and Rodent Models of Retinal Ganglion Cell Apoptosis

Purpose To investigate the protective effects of a novel cyclopeptide C*HSDGIC* (CHC) from the cyclization of Pituitary adenylate cyclase-activating polypeptide (PACAP) (1–5) in cellular and rodent models of retinal ganglion cell apoptosis. Methodology/Principal Findings Double-labeling immunohistochemistry was used to detect the expression of Thy-1 and PACAP receptor type 1 in a retinal ganglion cell line RGC-5. The apoptosis of RGC-5 cells was induced by 0.02 J/cm2 Ultraviolet B irradiation. MTT assay, flow cytometry, fluorescence microscopy were used to investigate the viability, the level of reactive oxygen species (ROS) and apoptosis of RGC-5 cells respectively. CHC attenuated apoptotic cell death induced by Ultraviolet B irradiation and inhibited the excessive generation of ROS. Moreover, CHC treatment resulted in decreased expression of Bax and concomitant increase of Bcl-2, as was revealed by western-blot analysis. The in vivo apoptosis of retinal ganglion cells was induced by injecting 50 mM N-methyl-D-aspartate (NMDA) (100 nmol in a 2 µL saline solution) intravitreally, and different dosages of CHC were administered. At day 7, rats in CHC+ NMDA-treated groups showed obvious aversion to light when compared to NMDA rats. Electroretinogram recordings revealed a marked decrease in the amplitudes of a-wave, b-wave, and photopic negative response due to NMDA damage. In retina receiving intravitreal NMDA and CHC co-treatment, these values were significantly increased. CHC treatment also resulted in less NMDA-induced cell loss and a decrease in the proportion of dUTP end-labeling-positive cells in ganglion cell line. Conclusions C*HSDGIC*, a novel cyclopeptide from PACAP (1–5) attenuates apoptosis in RGC-5 cells and inhibits NMDA-induced retinal neuronal death. The beneficial effects may occur via the mitochondria pathway. PACAP derivatives like CHC may serve as a promising candidate for neuroprotection in glaucoma.

[1]  A. Tamas,et al.  PACAP promotes neuron survival in early experimental diabetic retinopathy , 2014, Neurochemistry International.

[2]  N. Agarwal.  RGC-5 cells. , 2013, Investigative ophthalmology & visual science.

[3]  Akira Yoshikawa,et al.  PACAP Attenuates NMDA-Induced Retinal Damage in Association with Modulation of the Microglia/Macrophage Status into an Acquired Deactivation Subtype , 2013, Journal of Molecular Neuroscience.

[4]  P. Khaw,et al.  Neuroprotection and other novel therapies for glaucoma. , 2013, Current opinion in pharmacology.

[5]  R. Gallego-Pinazo,et al.  Evaluation of presumptive biomarkers of oxidative stress, immune response and apoptosis in primary open-angle glaucoma. , 2013, Current opinion in pharmacology.

[6]  Rong-jie Yu,et al.  The N-Terminal HSDCIF Motif Is Required for Cell Surface Trafficking and Dimerization of Family B G Protein Coupled Receptor PAC1 , 2012, PloS one.

[7]  F. Drago,et al.  Early changes in pituitary adenylate cyclase-activating peptide, vasoactive intestinal peptide and related receptors expression in retina of streptozotocin-induced diabetic rats , 2012, Peptides.

[8]  Rong-jie Yu,et al.  Effects of cyclopeptide C*HSDGIC* from the cyclization of PACAP (1–5) on the proliferation and UVB-induced apoptosis of the retinal ganglion cell line RGC-5 , 2012, Peptides.

[9]  J. Goldberg,et al.  Glaucoma 2.0: neuroprotection, neuroregeneration, neuroenhancement. , 2012, Ophthalmology.

[10]  A. Tamas,et al.  Protective effects of the neuropeptide PACAP in diabetic retinopathy , 2012, Cell and Tissue Research.

[11]  M. Guido,et al.  Expression of Novel Opsins and Intrinsic Light Responses in the Mammalian Retinal Ganglion Cell Line RGC-5. Presence of OPN5 in the Rat Retina , 2011, PloS one.

[12]  Takuya Fujisawa,et al.  Edaravone-loaded liposomes for retinal protection against oxidative stress-induced retinal damage. , 2011, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[13]  H. Takeuchi,et al.  Edaravone-loaded liposome eyedrops protect against light-induced retinal damage in mice. , 2011, Investigative ophthalmology & visual science.

[14]  Z. Tan,et al.  Hypoxia induces beta-amyloid in association with death of RGC-5 cells in culture. , 2011, Biochemical and biophysical research communications.

[15]  H. Hara,et al.  Crocetin prevents retinal degeneration induced by oxidative and endoplasmic reticulum stresses via inhibition of caspase activity. , 2011, European journal of pharmacology.

[16]  Eun Ha Lee,et al.  Isoquercitrin is the most effective antioxidant in the plant Thuja orientalis and able to counteract oxidative-induced damage to a transformed cell line (RGC-5 cells) , 2010, Neurochemistry International.

[17]  K. Kang,et al.  Sulbutiamine Counteracts Trophic Factor Deprivation Induced Apoptotic Cell Death in Transformed Retinal Ganglion Cells , 2010, Neurochemical Research.

[18]  F. Gallyas,et al.  Pituitary adenylate cyclase activating polypeptide in the retina: focus on the retinoprotective effects , 2010, Annals of the New York Academy of Sciences.

[19]  Juan Li,et al.  A novel cyclopeptide from the cyclization of PACAP(1–5) with potent activity towards PAC1 attenuates STZ-induced diabetes , 2010, Peptides.

[20]  R. K. Murthy,et al.  Evaluation of ultraviolet light toxicity on cultured retinal pigment epithelial and retinal ganglion cells , 2010, Clinical ophthalmology.

[21]  S. Shioda,et al.  Suppression of Rat Retinal Ganglion Cell Death by PACAP Following Transient Ischemia Induced by High Intraocular Pressure , 2010, Journal of Molecular Neuroscience.

[22]  A. Tamas,et al.  PACAP Improves Functional Outcome in Excitotoxic Retinal Lesion: An Electroretinographic Study , 2010, Journal of Molecular Neuroscience.

[23]  A. Tamas,et al.  Effects of PACAP in UV-A Radiation-Induced Retinal Degeneration Models in Rats , 2010, Journal of Molecular Neuroscience.

[24]  S. Shioda,et al.  Neuroprotective Effect of PACAP Against NMDA-Induced Retinal Damage in the Mouse , 2010, Journal of Molecular Neuroscience.

[25]  H. Vaudry,et al.  Pituitary Adenylate Cyclase-Activating Polypeptide and Its Receptors: 20 Years after the Discovery , 2009, Pharmacological Reviews.

[26]  H. Vaudry,et al.  Molecular and conformational determinants of pituitary adenylate cyclase-activating polypeptide (PACAP) for activation of the PAC1 receptor. , 2009, Journal of medicinal chemistry.

[27]  F. Corcho,et al.  Molecular dynamics (MD) simulations of VIP and PACAP27 , 2009, Biopolymers.

[28]  A. Tamas,et al.  Effects of Pituitary Adenylate Cyclase Activating Polypeptide and Its Fragments on Retinal Degeneration Induced by Neonatal Monosodium Glutamate Treatment , 2009, Annals of the New York Academy of Sciences.

[29]  Xun Xu,et al.  Erythropoietin protects retinal neurons and glial cells in early-stage streptozotocin-induced diabetic rats. , 2008, Experimental eye research.

[30]  A. Vingrys,et al.  Rodent electroretinography: Methods for extraction and interpretation of rod and cone responses , 2008, Progress in Retinal and Eye Research.

[31]  Shi-zhou Huang,et al.  The photopic negative response of flash ERG in nonproliferative diabetic retinopathy , 2008, Documenta Ophthalmologica.

[32]  N. Osborne.  Pathogenesis of ganglion "cell death" in glaucoma and neuroprotection: focus on ganglion cell axonal mitochondria. , 2008, Progress in brain research.

[33]  Y. Ueda,et al.  In vivo activation of N-methyl-d-aspartate receptors generates free radicals and reduces antioxidant ability in the rat hippocampus: Experimental protocol of in vivo ESR spectroscopy and microdialysis for redox status evaluation , 2007, Brain Research.

[34]  V. Batuman,et al.  Intravenous infusion of pituitary adenylate cyclase-activating polypeptide (PACAP) in a patient with multiple myeloma and myeloma kidney: A case study , 2007, Peptides.

[35]  I. Pang,et al.  Comparison of expression profile of neurotrophins and their receptors in primary and transformed rat retinal ganglion cells. , 2007, Molecular vision.

[36]  P. Marin,et al.  PACAP type I receptor transactivation is essential for IGF‐1 receptor signalling and antiapoptotic activity in neurons , 2007, The EMBO journal.

[37]  S. Orrenius.  Reactive Oxygen Species in Mitochondria-Mediated Cell Death , 2007, Drug metabolism reviews.

[38]  A. Arimura,et al.  Signaling cascades involved in neuroprotection by subpicomolar pituitary adenylate cyclase-activating polypeptide 38 , 2007, Journal of Molecular Neuroscience.

[39]  F. Gallyas,et al.  The neuroprotective effects of PACAP in monosodium glutamate-induced retinal lesion involve inhibition of proapoptotic signaling pathways , 2006, Regulatory Peptides.

[40]  N. Irwin,et al.  Pituitary adenylate cyclase-activating peptide (PACAP): Assessment of dipeptidyl peptidase IV degradation, insulin-releasing activity and antidiabetic potential , 2006, Peptides.

[41]  P. Ježek,et al.  Mitochondria in homeostasis of reactive oxygen species in cell, tissues, and organism. , 2005, The international journal of biochemistry & cell biology.

[42]  N. Cooper,et al.  Serum deprivation induces apoptotic cell death of transformed rat retinal ganglion cells via mitochondrial signaling pathways. , 2005, Investigative ophthalmology & visual science.

[43]  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.

[44]  N. Osborne,et al.  Retinal ischemia: mechanisms of damage and potential therapeutic strategies , 2004, Progress in Retinal and Eye Research.

[45]  S. Onoue,et al.  The neuropeptide PACAP attenuates β-amyloid (1–42)-induced toxicity in PC12 cells , 2002, Peptides.

[46]  S. Shioda,et al.  Gene Expression for PACAP Receptor mRNA in the Rat Retina by in Situ Hybridization and in Situ RT‐PCR , 2000, Annals of the New York Academy of Sciences.

[47]  M. Inatani,et al.  Neuroprotective effects of brain-derived neurotrophic factor in eyes with NMDA-induced neuronal death , 2000, Brain Research.

[48]  J. Martinou,et al.  Mitochondria as the central control point of apoptosis. , 2000, Trends in cell biology.

[49]  G. Aldini,et al.  Fluorescent probes as markers of oxidative stress in keratinocyte cell lines following UVB exposure. , 2000, Farmaco.

[50]  H. Vaudry,et al.  Pituitary adenylate cyclase-activating polypeptide and its receptors: from structure to functions. , 2000, Pharmacological reviews.

[51]  J C Reed,et al.  Bax directly induces release of cytochrome c from isolated mitochondria. , 1998, Proceedings of the National Academy of Sciences of the United States of America.