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(Investigative Ophthalmology and Visual Science. 2002;43:744-750.)
© 2002 by The Association for Research in Vision and Ophthalmology, Inc.

Inhibitory Effects of Pyrrolidine Dithiocarbamate on Endotoxin-Induced Uveitis in Lewis Rats

Kouichi Ohta1, Kohzo Nakayama2, Toru Kurokawa1, Takanobu Kikuchi1 and Nagahisa Yoshimura1

1 From the Departments of Ophthalmology and 2 Anatomy, Shinshu University School of Medicine, Matsumoto, Japan.

PURPOSE. To determine the effect of pyrrolidine dithiocarbamate (PDTC), an antioxidant nuclear factor (NF)-{kappa}B inhibitor, on the ocular inflammation induced by lipopolysaccharide (LPS).

METHODS. Endotoxin-induced uveitis (EIU) was produced by a footpad injection of 200 µg LPS in male Lewis rats. PDTC (200 mg/kg) was injected intraperitoneally 30 minutes before the LPS administration. The number of infiltrating cells and protein concentration in the aqueous humor (AqH) was determined from the AqH collected at 24 hours. Immunohistochemical staining with a monoclonal antibody against activated NF-{kappa}B was performed to evaluate the effect of PDTC on NF-{kappa}B activation. Interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-{alpha} mRNA expression in the iris-ciliary body (ICB) was determined by RNase protection assay (RPA). The levels of these cytokines and nitric oxide (NO) production were also determined.

RESULTS. The number of cells in the AqH was 1100 ± 254 cells/µL in rats injected with LPS and 90 ± 43 cells/µL in rats pretreated with PDTC (P < 0.001). The concentration of proteins was significantly lower in the AqH of rats pretreated with PDTC than in those without PDTC. The number of activated NF-{kappa}B–positive cells in the ICB was reduced by the PDTC treatment. The ICB at 6 hours after LPS injection exhibited increased expression of IL-1ß, IL-6, and TNF-{alpha} mRNAs, which was decreased after PDTC pretreatment. PDTC also significantly diminished the levels of these cytokines and nitrite-nitrate in the AqH.

CONCLUSIONS. These results suggest that PDTC reduces ocular inflammation in eyes with EIU by downregulating proinflammatory cytokine expression and by inhibiting the NF-{kappa}B–dependent signaling pathway.




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