Inhibitory effects of oral disulfiram on endotoxin-induced uveitis in rats.
OBJECTIVE
Disulfiram (DSF) exhibits a wide variety of biological activities, including an anti-inflammatory action, on which we focused our attention. The aim of the present study was to investigate the effect of oral DSF on endotoxin-induced uveitis (EIU) in rats.


METHODS
We investigated its effect upon cellular infiltration and protein leakage, as well as on the concentration of tumor necrosis factor-α (TNF-α), nitric oxide (NO), and prostaglandin E2 (PGE2) in the anterior chamber. Some eyes were enucleated for histologic examination and immunohistochemical analysis. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). One hour before the LPS injection, either 250, 500, or 750 mg/kg DSF was administered orally. Twenty-four hours later, the aqueous humor was collected from both eyes, and the number of infiltrating cells and protein concentration in the aqueous humor were determined. Levels of TNF-α, NO, and PGE2 were determined by enzyme-linked immunosorbent assay. Immunohistochemical analysis in the iris ciliary body (ICB) cells was performed to determine the expression of activated nuclear factor kappa B (NF-κB), inducible-nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2).


RESULTS
The oral administration with DSF suppressed, in a dose-dependent manner, the number of inflammatory cells, the protein concentration, and the levels of TNF-α, NO, and PGE2 in the aqueous humor and improved the histiologic status of the ocular tissue. The expression of activated NF-κB-positive cells in the ICB was significantly inhibited by oral administrated with DSF 3 hr after the LPS injection. The LPS-induced increased expressions of iNOS and COX-2 proteins in the ICB were also inhibited by oral DSF 24 hr after LPS injection.


CONCLUSIONS
The present results indicate that oral DSF suppresses the inflammation in EIU by inhibiting the NF-κB-dependent pathway and the subsequent production of pro-inflammatory mediators.
Kanai
Kazutaka
K
Department of Small Animal Internal Medicine, School of Veterinary Medicine, University of Kitasato, Towada, Aomori, Japan. kanai@vmas.kitasato-u.ac.jp
Itoh
Naoyuki
N
Yoshioka
Kazuki
K
Yonezawa
Tomohiro
T
Ikadai
Hiromi
H
Hori
Yasutomo
Y
Ito
Yoshimasa
Y
Nagai
Noriaki
N
Chikazawa
Seishirou
S
Hoshi
Fumio
F
Higuchi
Seiichi
S
D000284:Administration, Oral; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage*; D001082:Aqueous Humor / Q000187:drug effects / Q000378:metabolism*; D002924:Ciliary Body / Q000187:drug effects / Q000378:metabolism; D052246:Cyclooxygenase 2 Inhibitors / Q000008:administration & dosage; D015232:Dinoprostone / Q000037:antagonists & inhibitors; D004221:Disulfiram / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004334:Drug Administration Schedule; D004731:Endotoxins; D005136:Eye Proteins / Q000037:antagonists & inhibitors; D018836:Inflammation Mediators / Q000037:antagonists & inhibitors*; D008297:Male; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D009569:Nitric Oxide / Q000037:antagonists & inhibitors; D009994:Osmolar Concentration; D051381:Rats; D011917:Rats, Inbred Lew; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors; D014605:Uveitis / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology*
D000893:Anti-Inflammatory Agents; D052246:Cyclooxygenase 2 Inhibitors; D004731:Endotoxins; D005136:Eye Proteins; D018836:Inflammation Mediators; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D015232:Dinoprostone; D004221:Disulfiram