Effects of instillation of eyedrops containing disulfiram and hydroxypropyl-β-cyclodextrin inclusion complex on endotoxin-induced uveitis in rats.
OBJECTIVE
To investigate the anti-inflammatory effects of the instillation of disulfirum (DSF) eyedrops that enhance solubility using 2-hydroxypropyl-β-cyclodextrin (HPβCD) and hydroxypropylmethylcellulose (HPMC) on endotoxin-induced uveitis (EIU) in rats and mechanisms related to ocular inflammation.


METHODS
EIU was induced in male Lewis rats by subcutaneous injection of 200 μg lipopolysaccharide (LPS). DSF (0.125%, 0.25% and 0.5%) or commercially available 0.05% dexamethasone (Dexa) was topically applied to both eyes of rats 1 hour before, immediately after, and 1 and 2 hours after injection of LPS. The aqueous humor (AqH) was collected 24 hours after LPS injection, and the number of infiltrating cells, protein concentration, and levels of tumor necrosis factor-α (TNF-α), nitric oxide (NO) and prostaglandin E2 (PGE2) were determined. Immunohistochemical analysis of the iris ciliary body (ICB) cells was performed to determine the expression of activated nuclear factor κB (NF-κB), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2).


RESULTS
The topical administration with DSF suppressed, in a dose-dependent manner, the number of inflammatory cells, the protein concentration, and the levels of NO, TNF-α and PGE2 in the AqH and improved the histologic status of the ocular tissue. The anti-inflammatory potency of 0.5% DSF treatment was as strong as that of 0.05% Dexa. Topical DSF treatment also suppressed the activated NF-κB 3 hours after LPS injection, and iNOS and COX-2 expression in the ICB 24 hours after LPS injection.


CONCLUSIONS
The present results demonstrate that the topical instillation of DSF eyedrops suppresses the inflammation in EIU, suggesting a possible novel approach for the treatment of ocular inflammation.
Department of Small Animal Internal Medicine, School of Veterinary Medicine, University of Kitasato, Towada, Aomori, Japan. kanai@vmas.kitasato-u.ac.jp
Kazutaka
K
Kanai
Yoshimasa
Y
Ito
Noriaki
N
Nagai
Naoyuki
N
Itoh
Yasutomo
Y
Hori
Seishirou
S
Chikazawa
Fumio
F
Hoshi
Seiichi
S
Higuchi
D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage; D001082:Aqueous Humor / Q000378:metabolism; D003907:Dexamethasone / Q000008:administration & dosage; D015232:Dinoprostone / Q000378:metabolism; D004195:Disease Models, Animal; D004221:Disulfiram / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D008070:Lipopolysaccharides; D008297:Male; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D009883:Ophthalmic Solutions; D051381:Rats; D011917:Rats, Inbred Lew; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014605:Uveitis / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism; D047392:beta-Cyclodextrins / Q000008:administration & dosage*
D000893:Anti-Inflammatory Agents; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009883:Ophthalmic Solutions; D014409:Tumor Necrosis Factor-alpha; D047392:beta-Cyclodextrins; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D009569:Nitric Oxide; D003907:Dexamethasone; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D015232:Dinoprostone; D004221:Disulfiram