Reference - Detail
|Fukuda K, Ishida W, Miura Y, Kishimoto T, Fukushima A.
|Cytokine expression and barrier disruption in human corneal epithelial cells induced by alarmin released from necrotic cells.
|Jpn J Ophthalmol
PURPOSE:Dying cells release endogenous molecules known as alarmins that signal danger to surrounding tissue. We investigated the effects of necrotic cell-derived alarmins on cytokine expression and barrier function in human corneal epithelial cells.
METHODS:The release of interleukin (IL)-6 and IL-8 from immortalized human corneal epithelial (HCE) cells in culture was measured with enzyme-linked immunosorbent assays. The abundance of IL-6 and 8 mRNAs was quantitated by reverse transcription and real-time polymerase chain reaction analysis. Barrier function of HCE cells was evaluated by measurement of transepithelial electrical resistance (TER). The subcellular localization of the p65 subunit of the transcription factor NF-κB was determined by immunofluorescence analysis, and phosphorylation of the endogenous NF-κB inhibitor IκBα was examined by immunoblot analysis.
RESULTS:A necrotic cell supernatant prepared from HCE cells induced the up-regulation of IL-6 and 8 expression at both mRNA and protein levels as well as reduced TER in intact HCE cells. Among alarmins tested, only IL-1α (not IL-33 or HMGB1) mimicked these effects of the necrotic cell supernatant. Furthermore, IL-1 receptor antagonist (IL-1RA) and neutralizing antibodies to IL-1α (but not those to IL-1β) each attenuated the effects of the necrotic cell supernatant. Exposure of HCE cells to the necrotic cell supernatant also induced the phosphorylation and degradation of IκBα as well as translocation of the p65 subunit of NF-κB to the nucleus.
CONCLUSION:IL-1α released from necrotic corneal epithelial cells may trigger inflammatory responses at the ocular surface, including cytokine production and barrier disruption.
|Alarmins / adverse effects Cells, Cultured Corneal Diseases / chemically induced Corneal Diseases / genetics* Corneal Diseases / metabolism Cytokines / biosynthesis Cytokines / genetics Electric Impedance Enzyme-Linked Immunosorbent Assay Epithelium, Corneal / metabolism* Epithelium, Corneal / pathology Gene Expression Regulation* Humans Immunoblotting Interleukin-1alpha / biosynthesis Interleukin-1alpha / genetics* Interleukin-6 / biosynthesis Interleukin-6 / genetics* Interleukin-8 / biosynthesis Interleukin-8 / genetics* Necrosis / genetics Necrosis / metabolism Necrosis / pathology RNA / genetics* Reverse Transcriptase Polymerase Chain Reaction
|Human and Animal Cells