Suppressive effects of tranilast on eotaxin-1 production from cultured conjunctival fibroblasts.
OBJECTIVE
Eotaxin, a CC-chemokine with selective chemotactic effects for eosinophils, has been reported to play an important role in allergic conjunctival diseases. We previously reported that eotaxin is produced by conjunctival fibroblasts and keratocytes on stimulation with Th2 cytokines. Tranilast is known to have anti-allergic properties. In this study, we examined the inhibitory effect of tranilast, an anti-allergic drug, on eotaxin-1 production from cultured conjunctival fibroblasts.


METHODS
Conjunctival fibroblasts obtained from normal patients were cultured in DMEM/F12 medium. On the fifth passage, the cells were transferred to a 96-well plate and, after starvation for 24 hr, TNF-alpha, IL-4, and tranilast or dexamethasone were added. After 48 hr, the concentrations of eotaxin-1 in the supernatants were measured by ELISA, and the cells were tested for eotaxin-1 expression by real-time PCR.


RESULTS
Eotaxin-1 production was observed on simultaneous stimulation with TNF-alpha and IL-4. This production was inhibited by both tranilast and dexamethasone. Inhibition of eotaxin-1 expression was also observed by real-time PCR.


CONCLUSIONS
Eotaxin-1 production from conjunctival fibroblasts was inhibited by both tranilast and dexamethasone. These results suggest that the anti-allergic effect of tranilast may be partly due to the inhibition of eotaxin-1 production from conjunctival fibroblasts.
Hida
Richard Yudi
RY
Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
Takano
Yoji
Y
Okada
Naoko
N
Dogru
Murat
M
Satake
Yoshiyuki
Y
Fukagawa
Kazumi
K
Fujishima
Hiroshi
H
D018926:Anti-Allergic Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D054413:Chemokine CCL11 / Q000096:biosynthesis* / Q000235:genetics; D003228:Conjunctiva / Q000166:cytology*; D003907:Dexamethasone / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D062367:ortho-Aminobenzoates / Q000494:pharmacology*
D018926:Anti-Allergic Agents; D054413:Chemokine CCL11; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D062367:ortho-Aminobenzoates; D015847:Interleukin-4; D003907:Dexamethasone; C012293:tranilast