April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Activation of Nf-B P65 Protects Retinal Ganglion Cells After Retinal Ischemia-Reperfusion in Rat
Author Affiliations & Notes
  • J. Wang
    Glaucoma,
    Tianjin Medical University Eye Center, Tianjin, China
  • S. Jiang
    Glaucoma,
    Tianjin Medical University Eye Center, Tianjin, China
  • X. Li
    retina,
    Tianjin Medical University Eye Center, Tianjin, China
  • J. Ge
    Glaucoma, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
  • Footnotes
    Commercial Relationships  J. Wang, None; S. Jiang, None; X. Li, None; J. Ge, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 4148. doi:
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    • Get Citation

      J. Wang, S. Jiang, X. Li, J. Ge; Activation of Nf-B P65 Protects Retinal Ganglion Cells After Retinal Ischemia-Reperfusion in Rat. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4148.

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

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Abstract

Purpose: : To investigate whether NF-ΚB p65 is activated and the activation of NF-ΚB p65 protects retinal ganglion cells (RGCs) after retinal ischemia-reperfusion (I/R) in rat.

Methods: : Retinal ischemia was induced by transiently elevating the intraocular pressure. Animals were euthanized at different time points after the insult. Eyes were enucleated for semiquantitative real-time RT-PCR studies of NF-ΚB p65, IΚB- (endogenous inhibitor of NF-ΚB). Retinal paraffin sections were prepared for immunohistochemistry for phosphor-NF-ΚB p65, phosphor-IΚB-, and ED1 (a marker of microglial/phagocytic cells) antibodies. Two inhibitors of NF-ΚB, helenalin and carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG-132), and small interfering RNA (siRNA) of NF-ΚB p65 were given to animals to examine the expression of NF-ΚB p65 mRNA and the number of RGCs by cresyl violet staining and 3% Fluoro-Gold staining. The number of RGCs was counted.

Results: : Semiquantitative real-time RT-PCR showed elevated NF-ΚB p65 mRNA (n=6) levels at 2 to 12 hours. IΚB- mRNA (n=6) showed elevated levels at 8 hours, compared with that of normal (n=10). Immunoreactivity (IM) of phosphor-NF-ΚB p65 increased in the retinal ganglion cell layer, the inner plexiform layer and the inner nuclear layer showing at 8 and 12 hours respectively after I/R injury. NF-ΚB p65 and ED1 co-localized in microglia/macrophage-like cells. Phospho-IΚB- co-localized with ED1 in the same cells. Helenalin and MG-132 inhibited the expression of NF-ΚB p65 mRNA and decreased significantly the number of RGCs compared with that of the control (p<0.05). Down-expression of NF-ΚB p65 mRNA was detected using siRNA of NF-ΚB p65, which reduced significantly the number of RGCs compared with that of the vehicle and control (p<0.05).

Conclusions: : After retinal ischemia-reperfusion injury, NF-ΚB p65 is activated and the activation of NF-ΚB p65 in microglia/macrophage-like cells protects retinal ganglion cells in rat.

Keywords: ischemia • ganglion cells • transcription factors 
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