Effects of technetium-99 methylenediphosphonate on cytokine-induced activation of retro-ocular fibroblasts from patients with Graves' ophthalmopathy.
OBJECTIVE
To explore the effects of technetium-99 methylenediphosphonate (99Tc-MDP) on cell proliferation, hyaluronic acid (HA) synthesis and the expressions of human leucocyte antigen-DR (HLA-DR), intercellular adhesion molecule-1 (ICAM-1) on cultured retro-ocular fibroblasts (RFs) from patients with Graves' ophthalmopathy.


METHODS
After two to seven passages, cultured RFs were incubated for 72 h with interferon-γ (100 U/ml), interleukin-1 (100 U/ml) or tumour necrosis factor-α (100 U/ml) in the presence of 99Tc-MDP. Flow cytometry was used to investigate the expression of HLA-DR and ICAM-1. RF proliferation was assessed by 3H-thymidine incorporation assay. HA synthesis was measured by radioimmunoassay.


RESULTS
At base conditions, the percentage of positive cells of HLA-DR and ICAM-1 on RFs was 6.70±3.06% and 5.29±3.02%, respectively, and the synthesis of HA was 337.8±42.7 ng/ml. Compared with basal values, 72-h incubation with cytokine significantly enhanced the expression of HLA-DR and ICAM-1, and HA synthesis. 99Tc-MDP (1 ng/ml) had little effect on cytokine-induced HLA-DR and ICAM-1 expression, and HA synthesis. When the concentration ranged from 10 to 100 ng/ml, 99Tc-MDP inhibited cytokine-induced RF activation in a dose-dependent manner. 99Tc-MDP also inhibited the proliferation of RFs in a dose-dependent manner. It was also found that 99Tc-MDP had the same effect on cytokine-induced RFs and skin fibroblasts from patients with normal individual conditions.


CONCLUSIONS
99Tc-MDP could inhibit cytokine-induced activation of RFs derived from patients with Graves' ophthalmopathy.
Department of Endocrinology and Metabolism, The First Affiliated Hospital, Henan College of Traditional Chinese Medicine, Zhengzhou, People's Republic of China.
Shu-Xun
SX
Yan
Ying
Y
Wang
Gui-Jun
GJ
Peng
Xin-Ping
XP
Lu
Yu
Y
Fu
D000328:Adult; D016022:Case-Control Studies; D049109:Cell Proliferation / Q000187:drug effects; D016207:Cytokines / Q000494:pharmacology*; D005123:Eye / Q000473:pathology*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D005786:Gene Expression Regulation / Q000187:drug effects; D049970:Graves Ophthalmopathy / Q000378:metabolism / Q000473:pathology*; D006684:HLA-DR Antigens / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000096:biosynthesis; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008297:Male; D012867:Skin / Q000166:cytology; D013669:Technetium Tc 99m Medronate / Q000494:pharmacology*
D016207:Cytokines; D006684:HLA-DR Antigens; D018799:Intercellular Adhesion Molecule-1; D006820:Hyaluronic Acid; D013669:Technetium Tc 99m Medronate