| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
–Induced Apoptosis in Corneal Fibroblasts by Transcription Factor NF-
B
1 From the Department of Ophthalmology, University of Washington School of Medicine, Seattle; the 2 Department of Cell Biology and Eye Institute, The Cleveland Clinic Foundation, Ohio; and the 3 Department of Ophthalmology, Sungkyunkwan University School of Medicine, Seoul, South Korea.
PURPOSE. Previous studies have suggested no role for tumor necrosis factor
(TNF)-
in the modulation of apoptosis in corneal fibroblasts.
However, recent investigations have demonstrated that nuclear factor
(NF)-
B activation by TNF- mediates negative apoptotic effects
that must be blocked to unmask the apoptotic effects of TNF- in
vitro. The purpose of this study was to investigate the role of
transcription factor NF-B in the suppression of TNF-–induced
apoptosis of corneal fibroblasts.
METHODS. mRNA was detected by reverse transcription–polymerase chain reaction
(RT-PCR) and RNase protection assay. Proteins were detected by
immunocytochemistry and immunoprecipitation with Western blot analysis.
Cell death was evaluated by trypan blue exclusion assay in corneal
fibroblasts treated with TNF- in presence or absence of the specific
inhibitor of NF-B activation, SN50, actinomycin D, or actinomycin D
with dexamethasone, ketorolac tromethamine, or diclofenac sodium.
Apoptosis was monitored by trypan blue exclusion, colorimetric cell
assay, CPP32 activation assay, DNA fragmentation assay, and
transmission electron microscopy. NF-B activation was monitored
using electrophoretic gel shift assay.
RESULTS. TNF-, TNF receptor (R)I, and TNFRII mRNAs were detected in all three
cultured corneal cell types and in ex vivo corneal epithelium using
RT-PCR. TNF- mRNA was also detected in ex vivo corneal epithelium,
corneal epithelial cells, and stromal fibroblasts with the RNase
protection assay. TNF-, TNFRI, and TNFRII proteins were detected by
immunocytochemistry in all three major corneal cell types in human
corneal tissue. TNF- protein was also detected in ex vivo corneal
epithelium, primary corneal epithelial cells, and primary stromal
fibroblasts using immunoprecipitation and Western blot analysis.
TNF- stimulated corneal fibroblast cell death when NF-B
activation was blocked with actinomycin D or SN50. Enhanced cell death
was noted with dexamethasone, ketorolac tromethamine, or diclofenac
sodium when used in the presence, but not in the absence, of
actinomycin D. A gel shift assay revealed induction of NF-B by
TNF- and suppression of induction in the presence of actinomycin D
or SN50, but not by the control peptide SN50M.
CONCLUSIONS. The TNF- receptor system is expressed in the cornea, and NF-B
activation is an important regulator of TNF-–mediated corneal
fibroblast apoptosis. Nonsteroidal anti-inflammatory agents or
corticosteroids may potentiate corneal fibroblast apoptosis in response
to cytokine stimulation.
This article has been cited by other articles:
|
|
 |
|
  C. Meltendorf, G. J. Burbach, J. Buhren, R. Bug, C. Ohrloff, and T. Deller Corneal Femtosecond Laser Keratotomy Results in Isolated Stromal Injury and Favorable Wound-Healing Response Invest. Ophthalmol. Vis. Sci., May 1, 2007; 48(5): 2068 - 2075. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. He and H. E. P. Bazan Synergistic Effect of Platelet-Activating Factor and Tumor Necrosis Factor-{alpha} on Corneal Myofibroblast Apoptosis. Invest. Ophthalmol. Vis. Sci., March 1, 2006; 47(3): 883 - 891. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. S. Lakka, C. S. Gondi, D. H. Dinh, W. C. Olivero, M. Gujrati, V. H. Rao, C. Sioka, and J. S. Rao Specific Interference of Urokinase-type Plasminogen Activator Receptor and Matrix Metalloproteinase-9 Gene Expression Induced by Double-stranded RNA Results in Decreased Invasion, Tumor Growth, and Angiogenesis in Gliomas J. Biol. Chem., June 10, 2005; 280(23): 21882 - 21892. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. E. Wilson, R. R. Mohan, M. Netto, V. Perez, D. Possin, J. Huang, R. Kwon, A. Alekseev, and J. P. Rodriguez-Perez RANK, RANKL, OPG, and M-CSF Expression in Stromal Cells during Corneal Wound Healing Invest. Ophthalmol. Vis. Sci., July 1, 2004; 45(7): 2201 - 2211. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Rezai, L. Farrokh-Siar, E. M. Gasyna, J. T. Ernest, and G. A. van Seventer The Effect of Type I and II Interferons on Human Fetal Retinal Pigment Epithelium-Induced Apoptosis in Jurkat T Cells Invest. Ophthalmol. Vis. Sci., July 1, 2003; 44(7): 3130 - 3134. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. B. Lee, J. A. Javier, J.-H. Chang, C. C. Chen, T. Kato, and D. T. Azar Confocal and Electron Microscopic Studies of Laser Subepithelial Keratomileusis (LASEK) in the White Leghorn Chick Eye Arch Ophthalmol, December 1, 2002; 120(12): 1700 - 1706. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. D'Acquisto, M. J. May, and S. Ghosh Inhibition of Nuclear Factor Kappa B (NF-B):: An Emerging Theme in Anti-Inflammatory Therapies Mol. Interv., February 1, 2002; 2(1): 22 - 35. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-W. Hong, J. J. Liu, J.-S. Lee, R. R. Mohan, R. R. Mohan, D. J. Woods, Y.-G. He, and S. E. Wilson Proinflammatory Chemokine Induction in Keratocytes and Inflammatory Cell Infiltration into the Cornea Invest. Ophthalmol. Vis. Sci., November 1, 2001; 42(12): 2795 - 2803. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. E. Wilson, R. R. Mohan, J.-W. Hong, J.-S. Lee, R. Choi, and R. R. Mohan The Wound Healing Response After Laser In Situ Keratomileusis and Photorefractive Keratectomy: Elusive Control of Biological Variability and Effect on Custom Laser Vision Correction Arch Ophthalmol, June 1, 2001; 119(6): 889 - 896. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
