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Effects of PACAP on Intracellular Signaling Pathways in Human Retinal Pigment Epithelial Cells Exposed to Oxidative Stress

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Abstract

The integrity of retinal pigment epithelial cells is critical for photoreceptor survival and vision. Pituitary adenylate cyclase activating polypeptide (PACAP) exerts retinoprotective effects against several types of injuries in vivo, including optic nerve transection, retinal ischemia, excitotoxic injuries, UVA-induced lesion, and diabetic retinopathy. In a recent study, we have proven that PACAP is also protective in oxidative stress-induced injury in human pigment epithelial cells (ARPE-19 cells). The aim of the present study was to investigate the possible mechanisms of this protection. ARPE cells were exposed to a 24-h hydrogen peroxide treatment. Expressions of kinases and apoptotic markers were studied by complex array kits and Western blot. Oxidative stress induced the activation of several apoptotic markers, including Bad, Bax, HIF-1α, several heat shock proteins, TNF-related apoptosis-inducing ligand, and Fas-associated protein with death domain, while PACAP treatment decreased them. The changes in the expression of MAP kinases showed that PACAP activated the protective ERK1/2 and downstream CREB, and decreased the activation of the pro-apoptotic p38MAPK and c-Jun N-terminal kinase, an effect opposite to that observed with only oxidative stress. Furthermore, PACAP increased the activation of the protective Akt pathway. In addition, the effects of oxidative stress on several other signaling molecules were counteracted by PACAP treatment (Chk2, Yes, Lyn, paxillin, p53, PLC, STAT4, RSK). These play a role in cell death, cell cycle, inflammation, adhesion, differentiation and proliferation. In summary, PACAP, acting at several levels, influences the balance between pro- and anti-apoptotic factors in favor of anti-apoptosis, thereby providing protection in oxidative stress-induced injury of human retinal pigment epithelial cells.

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Acknowledgments

This work was supported by SROP 4.1.2.B-10/2/KONV-20/0-0002, SROP-4.2.2/B-10/1-2010-0029, OTKA K72592, 100144, CNK78480, Momentum-Program of the Hungarian Academy of Sciences, Bolyai Scholarship, Akira Arimura Foundation, and Richter Gedeon Centenary Foundation.

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Authors and Affiliations

  1. Department of Anatomy, PTE-MTA “Lendulet” PACAP Research Team, University of Pecs, 7624, Pecs, Szigeti u 12, Hungary

    E. Fabian, D. Reglodi & A. Tamas

  2. Department of Biochemistry and Medical Chemistry, University of Pecs, Pecs, Hungary

    L. Mester, A. Szabo & K. Kovacs

  3. Department of Experimental Zoology and Neurobiology, University of Pecs, Pecs, Hungary

    K. Szabadfi

  4. Department of Medical Chemistry, University of Szeged, Szeged, Hungary

    G. Toth

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  1. E. Fabian
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  2. D. Reglodi
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  3. L. Mester
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  4. A. Szabo
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  5. K. Szabadfi
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  8. K. Kovacs
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Correspondence to D. Reglodi.

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Fabian, E., Reglodi, D., Mester, L. et al. Effects of PACAP on Intracellular Signaling Pathways in Human Retinal Pigment Epithelial Cells Exposed to Oxidative Stress. J Mol Neurosci 48, 493–500 (2012). https://doi.org/10.1007/s12031-012-9812-7

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  • DOI: https://doi.org/10.1007/s12031-012-9812-7

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