Celecoxib and Pioglitazone as Potential Therapeutics for Regulating TGF-β-Induced Hyaluronan in Dysthyroid Myopathy.
OBJECTIVE
To investigate the role of extraocular muscles (EOM) myoblasts in Graves ophthalmopathy (GO) pathology and the effect of a cyclooxygenase (COX)-2 inhibitor and a peroxisome proliferator-activated receptor (PPAR)-γ agonist in its treatment.


METHODS
Myoblasts were isolated and cultured from EOM of 10 patients with GO and 4 without (non-GO). The cultured myoblasts were treated with IFN-γ, insulin-like growth factor (IGF)-1, IL-1β, and TNF-α, and the effect on PPAR-γ, COX-2, TGF-β, and thyroid stimulating hormone receptor (TSH-R) expressions were assessed using real-time (RT)-PCR, ELISA, and Western blot. The effect of a COX-2 inhibitor and a PPAR-γ agonist on the expression of TGF-β, hyaluronan synthases (HAS)-1, -2, and -3, and hyaluronan (HA) were further evaluated.


RESULTS
Real-time PCR showed significant upregulation in PPAR-γ, COX-2, TGF-β, and TSH-R mRNA expression in GO myoblasts when treated with TNF-α but not in the non-GO. While IFN-γ and IGF-1 had no significant effect, IL-1β did upregulate COX-2 expression. These results were further confirmed by ELISA and Western blotting. Tumor necrosis factor α-induced TGF-β in turn significantly increased HA expression and HAS3 level, but not HAS1 and HAS2. The cyclooxygenase 2 inhibitor and PPAR-γ agonist substantially diminished this TNF-α-induced TGF-β, HA, and HAS3 expression.


CONCLUSIONS
These results demonstrate the role of EOM myoblasts in the pathogenesis of GO. The cyclooxygenase 2 inhibitor and PPAR-γ agonist in this study are potential treatments for GO due to their ability to suppress TNF-α-induced TGF-β, HAS, and HA upregulation.
Ocular Surface Center, Miami, Florida, United States.
Anny M S
AM
Cheng
Florida International University Herbert Wertheim College of Medicine, Miami, Florida, United States.
Han Y
HY
Yin
School of Medicine, National Taiwan University, Taipei, Taiwan.
Angus
A
Chen
Institute of Molecular Medicine, National Taiwan University, Taipei, Taiwan.
Ya-Wen
YW
Liu
Institute of Molecular Medicine, National Taiwan University, Taipei, Taiwan.
Mei-Chun
MC
Chuang
Ocular Surface Center, Miami, Florida, United States.
Hua
H
He
Ocular Surface Center, Miami, Florida, United States.
Sean
S
Tighe
Ocular Surface Center, Miami, Florida, United States.
Hosam
H
Sheha
School of Medicine, National Taiwan University, Taipei, Taiwan 5Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan.
Shu-Lang
SL
Liao
D000328:Adult; D015153:Blotting, Western; D016022:Case-Control Studies; D000068579:Celecoxib / Q000627:therapeutic use*; D002478:Cells, Cultured; D052246:Cyclooxygenase 2 Inhibitors / Q000627:therapeutic use*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D049970:Graves Ophthalmopathy / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D006801:Humans; D006820:Hyaluronic Acid / Q000502:physiology*; D008297:Male; D008875:Middle Aged; D032446:Myoblasts / Q000187:drug effects / Q000502:physiology; D009801:Oculomotor Muscles / Q000166:cytology / Q000378:metabolism; D047495:PPAR gamma / Q000819:agonists*; D000077205:Pioglitazone; D060888:Real-Time Polymerase Chain Reaction; D045162:Thiazolidinediones / Q000627:therapeutic use*; D016212:Transforming Growth Factor beta / Q000502:physiology*
D052246:Cyclooxygenase 2 Inhibitors; D047495:PPAR gamma; D045162:Thiazolidinediones; D016212:Transforming Growth Factor beta; D006820:Hyaluronic Acid; D000068579:Celecoxib; D000077205:Pioglitazone