Effect of genistein on the angiogenesis-related factors secreted by fibroblast-like synoviocytes from rats with collagen Ⅱ induced arthritis / 中华风湿病学杂志

ABSTRACT

Objective To investigate the effect of genistein (gen) on the secretion of vascular endo-thelial growth factor (VEGF) and matrix metalloproteinases (MMPs-1, 2, 3, 9) by fibroblast-like synoviocytes (FLS) from rats with collagen Ⅱ induced arthritis (CIA). Methods The CIA was induced by collagen Ⅱ and complete Freund's adjuvant (CFA) into rats. The rats were scored based on the arthritis index(AI) once a week. At the sixth week, the X-ray of joints was taken. The synovial tissues from knee joints were examined pathologically. The primary fibroblast-like synowocytes were separated from the synovial tissue by collagenase digestion and cultured. Then the expression of VCAM-1 was estimated by flow cytometry. After adding gen (100, 200, 400 μmol/L) at different concentrations into the FLS, VEGF and MMP-1, 2, 3, and 9 of the supernatants were tested by indirect ELISA. Results After 3 days of type Ⅱ collagen and CFA injec-tion, the rats started to catch arthrocele and their arthritis index increased gradually. The arthrocele was most remarkable at the 3rd week. The AI, X-ray and pathological examination indicated that the model group were significantly different from the control group. After the synoviocytes were cultured to the 4th generation, the expression of VCAM-1 was as high as about 85.5%. It showed that most synoviocytes were changed to fibro-blast-like synoviocytes. Different concentrations of gen (100, 200, 400 μmol/L) added to FLS were compared and revealed that the VEGF and MMP-1, 2, 3, and 9 in the supematants were suppressed evidently and in a dose-dependent manner. Conclusion The CIA model can be successfully constructed by collagen type Ⅱ and CFA. Tthe primary FLS of rats' joint can be separated and cultured well by collagenase digestion. Certain levels of gen can suppress the secretion of VEGF and MMP-1, 2, 3 and 9 hy FLS. The affect is dose-depen-dent.