Effect Of Genistein On The Inflammatory And Angiogenesis-related Factors Secreted By Fibroblast-like Synoviocytes From Rats With Collagen Induced Arthritis

Objectives:To investigate the effect of genistein(gen) on IL-1β(interleukin-1β),TNF-α(tumor necrosis factor-α),VEGF(vascular endothelial growth factor) and MMPs(matrix metalloproteinases)(MMP-1,2,3,9) secreted by FLS(fibroblast-like synoviocytes) from rats with CIA(collagen induced arthritis).Subject:Primary fibroblast-like synoviocytes from rats with CIA.Methods:At first, the rats were scored with the arthritis index(AI)once a week after the CIA was induced with collagen typeⅡand complete Freund's adjuvant(CFA). At the sixth week, the X-ray of joints were taken out. After the rats were sacrificed, synovial tissue from knee joint were detected by pathological examination. The primary fibroblast-like synoviocytes were separated from the synovial tissue with collagenase digestion and cultured. The expression of VCAM-1 on synovial cell was estimated by flow cytometry.Finally, different concentration of gen(100μM, 200μM, 400μM) and methotrexate(MTX)(1mg.L-1) were given to the incubation of fibroblast-like synoviocytes. Indirect ELISA(enzyme linked immunosorbent assay) was used to measure the relative contents of IL-1β,TNF-α,VEGF and MMP-1,2,3,9 in the supernatants of FLS.Results:1,The rats with CIA were induced with collagen typeⅡand complete Freund's adjuvant. The rats caught arthrocele apparently three days later. After detected by the arthritis index(AI),X-ray and pathological detection, it indicated that the CIA was well construceted.2,After the primary synoviocytes were cultured to 4th generation, expression of VCAM-1 was tested up to 85.5%. It showed that most part of synoviocytes were fibroblast-like synoviocytes. It fits our experimental requirement.3,With the effect of different concentration of gen(100μM, 200μM, 400μM) and MTX(1mg.L-1)on FLS, the IL-1βand TNF-α, VEGF and MMP-1,2,3,9 in supernatants were apparently restrained and dose-dependent.Conclusions:1,The CIA model was successfully constructed, the primary FLS of rats'joint were separated and cultured well.2,Gen can suppress the secretion of TNF-αand IL-1βby FLS and is dose-dependent. Gen may improve the arthromeningitis of CIA rats by inhibitting the secretion of inflammatory factors.3,Gen can suppress the secretion of VEGF and MMP-1,2,3,9 in FLS, the supression is dose-dependent. Therefore, gen may resist the angiogenesis by inhibiting angiogenesis-related factors secreted by FLS.