Inhibitory effects of pyrrolidine dithiocarbamate on endotoxin-induced uveitis in Lewis rats.
OBJECTIVE
To determine the effect of pyrrolidine dithiocarbamate (PDTC), an antioxidant nuclear factor (NF)-kappaB inhibitor, on the ocular inflammation induced by lipopolysaccharide (LPS).


METHODS
Endotoxin-induced uveitis (EIU) was produced by a footpad injection of 200 microg 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-kappaB was performed to evaluate the effect of PDTC on NF-kappaB activation. Interleukin (IL)-1beta, 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/microL in rats injected with LPS and 90 +/- 43 cells/microL 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-kappaB-positive cells in the ICB was reduced by the PDTC treatment. The ICB at 6 hours after LPS injection exhibited increased expression of IL-1beta, 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-kappaB-dependent signaling pathway.
Ohta
Kouichi
K
Department of Ophthalmology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan. kohta@hsp.md.shinshu-u.ac.jp
Nakayama
Kohzo
K
Kurokawa
Toru
T
Kikuchi
Takanobu
T
Yoshimura
Nagahisa
N
D000818:Animals; D000975:Antioxidants / Q000627:therapeutic use*; D001082:Aqueous Humor / Q000166:cytology / Q000378:metabolism; D002924:Ciliary Body / Q000378:metabolism / Q000473:pathology; D015536:Down-Regulation; D019084:Fluorescent Antibody Technique, Indirect; D007375:Interleukin-1 / Q000235:genetics / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007498:Iris / Q000378:metabolism / Q000473:pathology; D008070:Lipopolysaccharides; D008297:Male; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D011759:Pyrrolidines / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D012475:Salmonella; D013859:Thiocarbamates / Q000627:therapeutic use*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D014606:Uveitis, Anterior / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology
D000975:Antioxidants; D007375:Interleukin-1; D015850:Interleukin-6; D008070:Lipopolysaccharides; D016328:NF-kappa B; D011759:Pyrrolidines; D012333:RNA, Messenger; D013859:Thiocarbamates; D014409:Tumor Necrosis Factor-alpha; C020972:pyrrolidine dithiocarbamic acid; D009569:Nitric Oxide