Inhibitory effect of Saposhnikovia divaricate polysaccharide on fibroblast-like synoviocytes from rheumatoid arthritis rat in vitro

ABSTRACT

To study the mechanism and inhibitory effect of Saposhnikovia divaricata polysaccharide [SDP] on fibroblast-like synoviocytes [FLS] isolated from rheumatoid arthritis rat model. Rheumatoid arthritis rat model was established by the classical composite factors including wind, cold, damp plus biological agents. The synovial tissues were digested with trypsin to isolate FLS cells. The different dosage of SDP was applied in culture. The cell viability was evaluated by MTT assay and the apoptosis was determined by analytic flow cytometry. The expression change of p53 gene was monitored by RT-PCR method. The production of secretory inflammation factors TNF-alpha and IL-1beta were determined by ELISA. The proliferative and apoptotic proteins such as Bcl-2, Bax, Caspase-3, MMP-1, MMP-3, P53 were measured by western blotting. Our data demonstrated that treatment with high concentration of SDP could enhance the expression of P53 at both mRNA [P<0.05] and protein [P<0.05] level, inhibit the secretion of TNF-alpha [P<0.05] and IL-1beta [P<0.05]. Simultaneously, the Bcl-2/Bax ratio and level of MMP-1, MMP-3 was significantly decreased, and apoptotic marker caspase-3 protein was increased. In addition, the FACS analysis consistently consolidated the apoptosis-inducing effect of SDP on RAFLS. SDP could significantly inhibit dysplasia of RAFLS via modulation of p53 expression and suppression of inflammatory factors, which suggested a potential therapeutic value for rheumatoid arthritis