The Influence Of P53in Synovial Inflammation

Objective. To investigate the role of p53on inflammation of fibroblast-like synoviocytes(FLS), including the relationship between p53and NF-kB, p38, TLRs and their downstream pro-inflammatory cytokines.Materials and Method. Synovial tissues were obtained from patients at joint replacement surgery, and FLS were cultured in vitro.(1)FLS was co-incubated with IL-17at different concentration respectively for24hours. The supernatant was then collected for ELISA assay to dctcet Cytokines, while the cells were lysed for Westem blot to evaluate the expression of p53.(2)P53small interfering RNA (siRNA) or scrambled siRNA (sc siRNA) were electrotransfected into FLS. Three days later step(1) was repeated, RNA from FLS was extrated and then TLRs were amplified by real-time PCR, or FLS was co-incubated with IL-17(40ng/ml) for20minutes and then lyzed for Western Blot to identify the activation of signal transduction pathways like NF-κB and p38.(3) Ad-p53or Ad-Lacz were transfected into FLS. One days later step(1) was repeated, or the cells were lysed for Western blot to evaluate the expression of p53.Results.(1)IL-17can inhibit expression of p53and stimulate FLS overexpressing IL-6(P<0.05)(2)P53siRNA can effectively knockdown p53, leading To significant increase of IL-6secretion (p<0.05).(3) When IL-17was introduced, suppression of p53by p53siRNA provokes activation of P-p38and P-p65, compared with control group.(4)Transfection of p53siRNA into FLS induces decline of TLR2and TLR3along with the suppression of p53.(5)Transfection of Ad-p53(MOI=100) into FLS increases the expression of P53.Conclusions. Inhibition of p53leads to activation of signal transduction pathways like NF-κB and p38, and low expression of TLRs, results in subsequent elevation of downstream expression of pro-inflammatory cytokines. Overexpression of p53can’t decrease expression of IL-6.