Effects of diallyl trisulfide in human fibroblast-like synoviocytes from rheumatoid arthritis

Abstract

Objective To study the proliferation and inflammatory phenotypes of human fibroblast-like synoviocytes (FLS) induced by tumor necrosis factor-α (TNF-α). Methods The rheumatoid arthritis (RA)-FLS were cultured in vitro, then treated with different concentrations of diallyl trisulfide(DATS). The proliferation activity was detected by CCK-8 method. Then TNF-α was used to stimulate the RA-FLS, mRNA and protein expression of interleukin (IL)-6, matrix metalloproteinases (MMP)-1 and vascular endothelial growth factor(VEGF) were detected by quantitative real time polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA). Differences among groups were determined by one-way analysis of variance (ANOVA), LSD-t test was used for comparison between 2 groups. Results RA-FLS was successfully isolated and cultured in vitro. The positive rate of CD90 and CD29 in the RA-FLS was more than 90%. The proli-feration activity of RA-FLS treated with 100 μmoL/L, 200 μmol/L and 300 μmol/L DATS was(98.92±0.40)%, (95.91±0.32)%, (94.05±0.24)%, respectively. As Compared with the normal control group, the pro-liferation activity of RA-FLS was lower, and the statistically significant difference is between normal control group 200 μmol/L and 300 μmol/L DATS (t=-4.46, P<0.05; t=-7.98, P<0.05). After TNF-α stimulation, the expression of IL-6's mRNA in experiment group (100 μmol/L DATS) is higher compared with the model control group (t=5.74, P<0.05), but the change of IL-6's protein is no significant difference(t=-0.49, P=0.627). The differences of mRNA and protein expression levels of MMP-1 and VEGF between the experiment group (100 μmol/L DATS) and the model control group were not statistically significant. The relative mRNA level[(0.42±0.06), t=-23.47, P<0.05; (0.14±0.039), t=-36.59, P<0.05; (0.36±0.09), t=-13.1, P<0.05)] and the protein levels [(108.0±4.7) ng/L, t=-63.79, P<0.05, (26.0±1.0) ng/L, t=-9.68, P<0.05; (57.9±0.7), t=-34.59, P<0.05] of IL-6, MMP-1, VEGF in experiment group (200 μmol/L DATS) were significantly decreased. And the relative mRNA level [(0.041±0.027), t=-38.48, P<0.05; (0.027±0.027), t=-41.22, P<0.05; (0.131±0.047), t=-17.74, P<0.05] and the protein levels [(24.2±2.3) ng/L, t=-88.69, P<0.05; (22.7±1.0) ng/L, t=-14.13, P<0.05; (34.5±1.7),t=-48.45, P<0.05] of IL-6, MMP-1, VEGF in the experiment group(300 μmol/L DATS) were also significantly decreased. The difference between the two groups was significant (t=-24.89, P<0.05; t=-4.45, P<0.05; t=-13.87, P<0.05). Conclusion DATS can inhibit the proliferation and the effect of TNF-α induced secretion of IL-6, MMP-1 and VEGF in RA-FLS. The effect is dose-dependent. Key words: Arthritis, rheumatoid; Fibroblast-like synoviocyt; Diallyl trisulfide