March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Expression of Inducible Nitric Oxide Synthase and Cationic Amino Acid Transporter (CAT) In Endotoxin-Induced Uveitis
Author Affiliations & Notes
  • Chang-Hao Yang
    Ophthalmology, National Taiwan Univ Hospital, Taipei, Taiwan
  • Yi-An Lee
    Ophthalmology, National Taiwan Univ Hospital, Taipei, Taiwan
  • Chung-May Yang
    Ophthalmology, National Taiwan Univ Hospital, Taipei, Taiwan
  • Muh-Shy Chen
    Ophthalmology, National Taiwan Univ Hospital, Taipei, Taiwan
  • Footnotes
    Commercial Relationships  Chang-Hao Yang, None; Yi-An Lee, None; Chung-May Yang, None; Muh-Shy Chen, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 1244. doi:
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      Chang-Hao Yang, Yi-An Lee, Chung-May Yang, Muh-Shy Chen; Expression of Inducible Nitric Oxide Synthase and Cationic Amino Acid Transporter (CAT) In Endotoxin-Induced Uveitis. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1244.

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

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Abstract

Purpose: : Nitric oxide (NO) plays a significant role in the pathogenesis of ocular inflammation. The synthesis of NO depends on cell membrane protein cationic amino acid transporter (CAT) to transfer arginine into the cell and inducible nitric oxide synthase (iNOS) to convert arginine into NO. The purpose of this study is to examine the expression of CAT and iNOS in endotoxin-induced uveitis (EIU).

Methods: : EIU was induced by a footpad injection of 200 μg LPS in Lewis rats. Proteasome inhibitor bortezomib (valcade) was injected intraperitoneally 30 minutes before the LPS administration. The rats were sacrificed 24 hours later and the eyes were enucleated. The expression of iNOS and CAT mRNA and protein in the iris and ciliary body was determined by RT-PCT and Western blotting. The production of NO in aqueous was measured by ELISA. Immunochemical staining of the iris and ciliary body was performed to evaluate the inhibitory effect of valcade. . The interaction of NF-ΚB activation, CAT, and iNOS induction was assessed by electrophoretic mobility shift assay (EMSA).

Results: : Proteasome inhibitor valcade abrogated the inflammation of EIU as demonstrated by immunochemical stain and the decreased NO production in aqueous. The expression of iNOS and CAT mRNA and protein in EIU mouse eyes were suppressed by valcade. EMSA revealed that NF-ΚB activation was responsible for iNOS and CAT induction.

Conclusions: : NF-ΚB is an essential transcription factor not only for the induction of iNOS, but also for the up-regulation of CAT. The simultaneous up-regulation of CAT and iNOS may play an important role in the pathogenesis of EIU.

Keywords: uveitis-clinical/animal model • inflammation • immunomodulation/immunoregulation 
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