Open AccessThe expression of inducible nitric oxide synthase in human retinal pigment epithelial cells under stimulation of proinflammatory cytokine tumor necrosis factor-α
Author(s) -
I-Mo Fang,
Chang-Hao Yang,
Chung-May Yang,
Muh-Shy Chen
Publication year - 2012
Publication title -
taiwan journal of ophthalmology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.519
H-Index - 9
eISSN - 2211-5072
pISSN - 2211-5056
DOI - 10.1016/j.tjo.2011.11.001
Subject(s) - lactacystin , nitric oxide synthase , tumor necrosis factor alpha , proinflammatory cytokine , mg132 , nitric oxide , microbiology and biotechnology , western blot , cytokine , proteasome inhibitor , biology , medicine , chemistry , immunology , proteasome , endocrinology , inflammation , biochemistry , gene
PurposeTo elucidate the effects of tumor necrosis factor (TNF)-α on the expression of inducible nitric oxide synthase (iNOS) in retinal pigment epithelial cells in vitro.MethodsARPE-19 cell lines were cultured with TNF-α stimulation, and then treated with proteasome inhibitors (MG132 or lactacystin) for 30 minutes. The expression of iNOS was determined by RT-PCR and Western blot. The expression of nitric oxide (NO) was determined by an enzyme-linked immunosorbent assay. The interaction of nuclear factor kappa-B (NF-κB) activation and iNOS induction was assessed by electrophoretic mobility shift assay.ResultsThe expression of iNOS in ARPE-19 was induced by TNF-α in a dose-dependent manner. Upregulation of iNOS resulted in increased production of NO. iNOS induced by TNF-α could be inhibited by MG-132 and lactacystin. Supershift assay revealed that NF-κB activation was responsible for iNOS induction.ConclusionTNF-α could induce iNOS expression and NO production in RPE cells, at least in part, via an NF-κB signal pathway