May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Anti-Oxidative Capacity of Thymosin Beta-4 in Human Cornea Epithelial Cells
Author Affiliations & Notes
  • J. H. Ho
    Dept of Ophthalmology, Buddist Tzu Chi General Hospital-Taipei, Taipei, Taiwan
    Institute of Biopharmaceutical Science, Department of Medical Research & Education,
    National Yang-Ming University, Taipei, Taiwan
  • K.-C. Tseng
    Institute of Biopharmaceutical Science, Department of Medical Research & Education,
    Taipei Veterans General Hospital, Taipei, Taiwan
  • W.-H. Ma
    Institute of Biopharmaceutical Science, Department of Medical Research & Education,
    Taipei Veterans General Hospital, Taipei, Taiwan
  • K.-H. Chen
    Institute of Clinical Medicine, Department of Ophthalmology,
    Taipei Veterans General Hospital, Taipei, Taiwan
  • O. K. Lee
    Institute of Clinical Medicine, Department of Ophthalmology,
    National Yang-Ming University, Taipei, Taiwan
  • Y. Su
    Institute of Biopharmaceutical Science, Department of Medical Research & Education,
    National Yang-Ming University, Taipei, Taiwan
  • Footnotes
    Commercial Relationships  J.H. Ho, None; K. Tseng, None; W. Ma, None; K. Chen, None; O.K. Lee, None; Y. Su, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 2958. doi:
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      J. H. Ho, K.-C. Tseng, W.-H. Ma, K.-H. Chen, O. K. Lee, Y. Su; Anti-Oxidative Capacity of Thymosin Beta-4 in Human Cornea Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):2958.

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

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Abstract

Purpose: : We previously demonstrated that internalization was essential for exogenous thymosin beta-4 (Tβ4) to protect human cornea epithelial (HCE-T) cells against ROS-induced apoptosis, and the purpose of this study is to further elucidate its protective mechanism.

Methods: : HCE-T cells were incubated with histidine-tagged Tβ4 (1 µg/ml) for two hours before being treated with H2O2 (200 µM). Activation of caspases-8 and -9 was examined by flow cytometry and the latter was further analyzed by colorimetric assay. Intracellular reactive oxygen species (ROS) level was measured by flowcytometry using 2’,7’-dichlorodihydrofluorescein diacetate (H2DCFH-DA) as probes. Protein levels and gene expression of manganese-superoxide dismutase (Mn-SOD), copper/zinc-superoxide dismutase (Cu/Zn-SOD) and catalase were analyzed by Western blotting and real-time RT-PCR, respectively.

Results: : It was found that Tβ4 protected HCE-T cells from H2O2-induced apoptosis by reducing both caspase-9 activation and intracellular ROS levels. It was further discovered that Tβ4 increased the expression of Mn-SOD and Cu/Zn-SOD. Moreover, after the addition of H2O2, catalase was also up-regulated by Tβ4 at both transcription and translation levels.

Conclusions: : 4 was found capable of up-regulating anti-oxidative enzymes in human corneal epithelial cells and increasing the resistance of the human corneal epithelial cells to ROS-induced apoptosis.

Keywords: oxidation/oxidative or free radical damage • cornea: epithelium • antioxidants 
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