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Inhibitory effects of Triptolide on interferon‐γ‐induced human leucocyte antigen‐DR, intercellular adhesion molecule‐1, CD40 expression on retro‐ocular fibroblasts derived from patients with Graves’ ophthalmopathy
Author(s) -
Yan Shuxun,
Wang Ying
Publication year - 2006
Publication title -
clinical and experimental ophthalmology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.3
H-Index - 74
eISSN - 1442-9071
pISSN - 1442-6404
DOI - 10.1111/j.1442-9071.2006.01190.x
Subject(s) - triptolide , medicine , viability assay , flow cytometry , microbiology and biotechnology , antigen , immunology , in vitro , chemistry , biochemistry , biology , apoptosis
A bstract Purpose: To explore the effects of Triptolide, the principal active diterpenoid from the Chinese Medicinal Herb Tripterygium Wilfordii Hook F that has immunosuppressive and anti‐inflammatory properties, on cell proliferation, hyaluronic acid (HA) synthesis, and the expressions of human leucocyte antigen‐DR (HLA‐DR), intercellular adhesion molecule‐1 (ICAM‐1) and CD40 on cultured retro‐ocular fibroblasts (RFs) from patients with Graves’ ophthalmopathy. Methods: After two to five passages, cultured RFs were incubated for 48 h within a medium alone or in the presence of recombinant human interferon‐γ (IFN‐γ) and various concentrations of Triptolide. Cell viability was assessed by MTT (3‐[4.5‐dimethylahiazol‐2‐yl]‐2,5‐diphenyltetrazolium Bromide). RFs proliferation was assessed by [ 3 H]‐thymidine incorporation assay. Flow cytometry was used to investigate the amount of HLA‐DR, ICAM‐1 and CD40. HA synthesis was measured by radioimmunoassay. Results: Cell viability was not detrimentally affected when incubated with Triptolide from 0.01 µg/L to 10 µg/L for 48 h, and decreased with 20 µg/L Triptolide. The incorporation of [ 3 H]‐thymidine of RFs was 55 476 ± 15 842 cpm incubated with medium alone or 18 352 ± 3568 cpm with 10 µg/L Triptolide ( t = 5.600, P < 0.01). Initially, the percentage of positive cells of HLA‐DR, ICAM‐1 and CD40 on RFs were 4.75 ± 2.13%, 17.53 ± 10.12% and 6.38 ± 2.23%, respectively, and the synthesis of HA was 100 ± 12%. Compared with basal values, 48‐h incubation with IFN‐γ (100 U/mL) significantly enhanced the amount of HLA‐DR, ICAM‐1 and CD40, and HA synthesis. The values were 60.58 ± 10.12% ( t = 13.224, P < 0.01), 62.66 ± 18.17% ( t = 5.315, P < 0.01), 57.67 ± 13.61% ( t = 9.110, P < 0.01) and 164 ± 22% ( t = 9.238, P < 0.01), respectively. Triptolide 0.01 µg/L had little effect on IFN‐γ‐induced HLA‐DR, ICAM‐1 and CD40 amounts, as well as HA synthesis. When the concentration ranged from 0.1 µg/L to 10 µg/L, Triptolide inhibited IFN‐γ‐induced RFs activation in a dose‐dependent manner. It was also found that Triptolide had the same inhibiting effects on IFN‐γ‐induced RFs and skin fibroblasts from patients with normal individual conditions. Conclusions: Triptolide could inhibit IFN‐γ‐induced activation of RFs derived from patients with Graves’ ophthalmopathy.