Synergistic effect of platelet-activating factor and tumor necrosis factor-alpha on corneal myofibroblast apoptosis

Investigative Ophthalmology & Visual Science
Jiucheng He, Haydee E P Bazan

Abstract

Elimination of myofibroblasts after repair of corneal injury is essential for the maintenance of corneal transparency. In the current study, the role of platelet-activating factor (PAF) in combination with tumor necrosis factor (TNF)-alpha in corneal myofibroblast apoptosis was explored. Porcine corneal myofibroblasts (PCMs) were obtained from subcultured fibroblasts plated at a low density (5 cells/mm2). Mouse anti-alpha-smooth muscle actin antibody was used to identify the cell phenotype. Immunofluorescence was performed to localize PAF and TNF-alpha receptors in those cells. The reactivity of the antibodies was characterized by Western blot analysis. To induce myofibroblast apoptosis, PCMs were treated for 24 to 72 hours with methylcarbamyl-PAF (cPAF, 300 nM), a nonhydrolyzable PAF analogue, TNF-alpha (20 ng/mL), and TNF-alpha+cPAF, with or without LAU-0901 (150 nM), a novel PAF antagonist. Apoptosis was assayed by Hoechst 33258 and TUNEL staining and DNA laddering. 6-Diamidino-2-phenylindole (DAPI) was used for nuclear counterstaining. Images were recorded by fluorescence microscope. Immunofluorescence with a PAF-receptor (N terminus) polyclonal antibody showed that the receptor was expressed in both plasma and nuclear memb...Continue Reading

Citations

May 1, 2010·Molecular Neurobiology·Salomon Esquenazi, Haydee E P Bazan
Mar 2, 2010·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Najam A Sharif, Terry K Wiernas
Dec 8, 2009·Journal of Lipid Research·Sachidananda Kenchegowda, Haydee E P Bazan
Aug 29, 2012·Expert Opinion on Therapeutic Targets·Eirini NitodaMichael Koutsilieris
Mar 6, 2010·Prostaglandins, Leukotrienes, and Essential Fatty Acids·Jiucheng He, Haydee E P Bazan
Jun 13, 2006·American Journal of Physiology. Lung Cellular and Molecular Physiology·Carlos RamosMoisés Selman
Nov 1, 2020·The Ocular Surface·Sabeeh Kamil, Rajiv R Mohan
Dec 29, 2020·Prostaglandins & Other Lipid Mediators·Barbara DalmasoCarolina Beltrame Del Debbio

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis