May 2008
Volume 49, Issue 13
ARVO Annual Meeting Abstract  |   May 2008
Lipid Rafts and Actin Are Important for Thymosin Beta 4- Mediated Anti-Inflammatory Effects in Cornea
Author Affiliations & Notes
  • G. Sosne
    Wayne State Univ Sch of Med, Detroit, Michigan
    Ophthalmology and Anatomy & Cell Biology,
  • P. Qiu
    Wayne State Univ Sch of Med, Detroit, Michigan
  • M. Kurpakus-Wheater
    Wayne State Univ Sch of Med, Detroit, Michigan
  • Footnotes
    Commercial Relationships  G. Sosne, RegeneRx Biopharmaceuticals, C; P. Qiu, None; M. Kurpakus-Wheater, None.
  • Footnotes
    Support  Midwest Eye Banks
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3397. doi:
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      G. Sosne, P. Qiu, M. Kurpakus-Wheater; Lipid Rafts and Actin Are Important for Thymosin Beta 4- Mediated Anti-Inflammatory Effects in Cornea. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3397. doi:

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: : To investigate the relationship of Tβ4, actin/cytoskeleton dynamics, and membrane raft function in TNF-α signaling and inflammation.

Methods: : Human corneal epithelial cells and fibroblasts were pre-treated or not with Tβ4 (1 µg/ml) for one hour prior to treatment with cyotchalasin D (cytoD, 5 µM) or jasplakinolide (JP, 1 µM)(actin/cytoskeleton disrupters). Additional experiments also included treatment with the membrane raft inhibitor MβCD. The cells were then stimulated with TNF-α (10 ng/ml). Cell culture supernatants were assayed for IL-8 using ELISA and the cells were assayed for the effects on actin stress fibers (phalloidin staining) and NFΚB nuclear translocation and binding activity.

Results: : Disrupting the actin cytoskeleton with Tβ4, cytoD or with JP resulted in a significant inhibition of TNF-α-stimulated IL-8 secretion (P < 0.0001 for all comparisons). Disrupting membrane rafts before stimulating cells with TNF-α also significantly inhibited the secretion of IL-8 (** P < 0.0001). Treatment of the cells with MβCD caused a significant disruption of the actin cytoskeleton and decreased NFΚB nuclear translocation. In addition, Tβ4 treatment also disrupted TNF-α-stimulated actin stress fiber formation.

Conclusions: : Taken together, these results provide support for our hypothesis that the actin cytoskeleton interacts with membrane rafts to play a central role in TNF-α-mediated signaling involving the activation and translocation of NFΚB and resulting in IL-8 production. Along with our other studies demonstrating that Tβ4 suppresses TNF-α-mediated IL-8 production and NFΚB activation, these findings show that Tβ4 treatment also disrupts TNF-α-induced actin cytoskeletal changes and provide a basis for our hypothesis that Tβ4’s inhibitory effects on TNF-α-mediated inflammation may be related to the actin cytoskeleton changes seen.

Keywords: cornea: epithelium • signal transduction • cytokines/chemokines 

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