Mitochondrial Permeability Transition Pore in Inflammatory Apoptosis of Human Conjunctival Epithelial Cells and T Cells: Effect of Cyclosporin A
| Autor: | Larry A. Wheeler, Wanju Lee, Virginia L. Calder, Reuben Sana, Margarita Calonge, Michael E. Stern, Jianping Gao |
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| Rok vydání: | 2013 |
| Předmět: |
Programmed cell death
Fas Ligand Protein T-Lymphocytes Apoptosis Mitochondrial Membrane Transport Proteins Jurkat cells Conjunctival Diseases Fas ligand Interferon-gamma Jurkat Cells Annexin Cyclosporin a Humans fas Receptor Cells Cultured Caspase Inflammation Membrane Potential Mitochondrial Cell Death biology Mitochondrial Permeability Transition Pore Tumor Necrosis Factor-alpha Epithelial Cells Molecular biology Mitochondrial permeability transition pore Caspases Immunology Cyclosporine biology.protein Conjunctiva Immunosuppressive Agents |
| Zdroj: | Investigative Opthalmology & Visual Science. 54:4717 |
| ISSN: | 1552-5783 |
| DOI: | 10.1167/iovs.13-11681 |
| Popis: | Purpose To investigate the role of mitochondrial permeability transition pore (MPTP) and effect of cyclosporin A (CsA) on inflammatory apoptosis of human conjunctival epithelial cells (IOBA-NHC) and T cells. Methods IOBA-NHC and Jurkat cells were stimulated with IFNγ, TNFα, αFas, or PMA/αCD3, in the presence or absence of CsA. MPTP was determined using the calcein-cobalt technique. Mitochondrial membrane potential (ΔΨm) was measured with JC-1. Apoptosis was quantified by Annexin V/PI staining. Apoptosis mediators were evaluated by flow cytometry or Western blot. Results In IOBA-NHC, TNFα, and IFNγ induced MPTP opening, ΔΨm loss, and increased cell apoptosis. This was accompanied by upregulation of Fas/FasL; Bax; and caspase-3, -8, and -9 activation. Addition of CsA prevented IOBA-NHC from cell death by blocking MPTP opening, ΔΨm loss, Fas/FasL, and caspase activation. In PMA/αCD3-activated Jurkat T cells, MPTP opening and ΔΨm loss were increased along with cell apoptosis and upregulated Fas/FasL/caspase expressions. CsA further promoted T-cell apoptosis, ΔΨm loss, and upregulation of Fas/FasL/caspase. Conclusions Inflammation induces aberrant MPTP opening, resulting in an increased apoptosis in conjunctival epithelial cells. CsA protected IOBA-NHC from cell death by blocking both intrinsic and extrinsic apoptosis pathways. CsA promoted T-cell apoptosis via upregulating Fas/FasL and caspase activities with a minimal effect on MPTP. The findings suggest that the differential effect of CsA on T cells versus ocular surface resident epithelial cells may contribute to its therapeutic efficacy in treating ocular inflammation such as dry eye disease. |
| Databáze: | OpenAIRE |
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