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Hsuan-Yeh Pan, Mallika Valapala; Role of nuclear factor activated in T cells (NFAT) signaling pathway in the Retinal Pigment Epithelial cells. Invest. Ophthalmol. Vis. Sci. 2019;60(9):1941.
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The nuclear factor activated in T cells (NFAT) is a family of transpiration factors that play an important role in integrating Ca2+ signaling with a variety of inflammatory processes. Of the five identified NFAT family members (NFAT1-5), four isoforms NFAT1-4 are subject to dephosphorylation and activation by calcineurin. Dephosphorylation by calcineurin results in NFAT nuclear translocation and transcriptional induction of NFAT-regulated genes, including, interleukin (IL)-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, IFN-γ, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF). We investigated the role of NFAT proteins in activating transcription of inflammatory cytokines in the RPE.
Transcriptional induction of NFAT was measured using the adenoviral NFAT reporter vector (AD-NFAT-luc). NFAT transcriptional activity was inhibited by a peptide, MAGPHPVIVITGPHEE (VIVIT), which known to selectively prevent the interaction of NFAT with calcineurin and inhibit nuclear translocation of NFAT. Quantitative Real time-PCR (qRT-PCR) analysis was used to determine the expression of cytokines: Interleukin 6 (IL-6), Interleukin 8 (IL-8)
ARPE-19 cells transfected with the AD-NFAT-luc vector for 24 hrs followed by treatment with 10 µg/ml LPS for 24 hrs showed a significant induction in luciferase activity. LPS-induced NFAT luciferase activity was reduced by treatment of cells with the NFAT inhibitor, VIVIT for 6 hrs. qRT-PCR analysis revealed increased expression of IL-6 and IL-8 in cells treated with LPS compared to untreated cells. LPS-induced expression of IL-6 and IL-8 was inhibited by treatment with the NFAT inhibitor,VIVIT.
These results suggest an important role of NFAT in regulating the expression of IL-6 and IL-8 in the RPE. Furthermore, inhibition of NFAT decreases LPS- induced expression of inflammatory cytokines in the RPE.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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