Abstract
Purpose: To evaluate keratocyte viability and proinflammatory cytokine secretion induced by HSV-1 infection. Methods: Keratocytes were separated from corneal tissues obtained with the SMILE procedure, and an in vitro system was established to study HSV-1 infection in human keratocytes. Cell viability, HSV-1 genomic DNA copy number, and the expression levels of α-SMA, ALDH1A1, phospho-p38, p38, phospho-IRF3, and IRF3 were evaluated. Antibody array and ELISA kits were used to measure the production of proinflammatory cytokines and chemokines. Results: We found that HSV-1 infection reduced cell viability and activated keratocyte transdifferentiation into corneal fibroblast-like cells. Furthermore, p38 inhibition improved cell viability and IFN-β production and played an anti-inflammatory role by reducing the production of proinflammatory cytokines and chemokines. Conclusions: Our study reveals an important role played by keratocytes during innate immune responses and identifies p38 inhibition as a potential therapeutic approach to control ocular HSV-1 infection.
Keywords:
HSV-1; IFN-β; Keratocytes; inflammation; p38 inhibition.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Actins / genetics
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Actins / metabolism
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Aldehyde Dehydrogenase 1 Family / genetics
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Aldehyde Dehydrogenase 1 Family / metabolism
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Animals
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Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
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Blotting, Western
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Cell Survival
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Cells, Cultured
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Chlorocebus aethiops
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Corneal Keratocytes / drug effects*
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Corneal Keratocytes / virology
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Cytokines / metabolism
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DNA Copy Number Variations
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Dose-Response Relationship, Drug
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Enzyme-Linked Immunosorbent Assay
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Herpes Simplex / metabolism
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Herpes Simplex / prevention & control*
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Herpes Simplex / virology
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Herpesvirus 1, Human / physiology*
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Humans
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Imidazoles / pharmacology*
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Interferon Regulatory Factor-3 / genetics
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Interferon Regulatory Factor-3 / metabolism
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Microscopy, Fluorescence
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Pyridines / pharmacology*
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Real-Time Polymerase Chain Reaction
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Retinal Dehydrogenase / genetics
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Retinal Dehydrogenase / metabolism
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Time Factors
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Vero Cells
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p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors*
Substances
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ACTA2 protein, human
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Actins
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Anti-Inflammatory Agents, Non-Steroidal
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Cytokines
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IRF3 protein, human
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Imidazoles
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Interferon Regulatory Factor-3
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Pyridines
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Aldehyde Dehydrogenase 1 Family
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ALDH1A1 protein, human
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Retinal Dehydrogenase
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p38 Mitogen-Activated Protein Kinases
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SB 203580