Regulator Role And Mechanisms Of NLRC5 In The Cytokin And Itself Expression In AA Mice Synoviocyte

Rheumatoid arthritis’s main features are progressive synovitis and joint structural damage. Fibroblast-like synovial cells (FLS) plays a leading role in the development of RA, which exhibit abnormal proliferation and invasion status and lead to the destruction of joints and cartilage. In this process, IL-1β, TNF-a, IL-6 is considered to be an important factor leading to the activation of FLS. These inflammatory cytokines regulates gene expression, cell proliferation and migration activation process via different signaling pathways. Inflammatory process are interaction net of a variety of inflammatory cytokines.NLRC5 was discovered as a newly NOD-like receptor family member, which can inhibit the immune and inflammatory responses and may become a future target for the effective treatment of immune-related diseases. Nuclear transcription factor (NF-kB) plays a decisive role in numerous inflammatory reaction due to effect the release of inflammatory cytokine and cell proliferation and activation. Especially for the regulation of IL-1(3, TNF-a, IL-6 and other inflammatory cytokines in the development and progression of RA. Recent studies show that NLRC5 strongly inhibit NF-kB activation, which does this by inhibiting the phosphorylation and activation of IKK and IKKβ to prevent the subunit of NF-kB translocate to the nucleus, thereby inhibiting the expression of inflammatory cytokines and the inflammatory response. In addition, NLRC5 also inhibit I type interferon response through different mechanisms. This study observed NLRC5 expression in AA model FLSs mice, and discussed the relationship between NLRC5 and inflammatory cytokines IL-1β,TNF-α, IL-6, in search the intervention of the new therapeutic targets of RA. The main contents are summarized as follows:We choose adjuvant arthritis model which pathological features are similar with RA. And we extracted synovial cells for the vitro study. We constructed AA model using Freund’s complete adjuvant. we evaluated AA mice through local joint visually change and the secondary mouse paw swelling, joint histopathology staining and determination of serum concentrations of inflammatory cytokines. Then the highly expression of NLRC5 were observed in model synovial tissues and primary culture FLSs at different time points. And we used chemical analysis methods and western blot to detect the NLRC5 expression level in the synovial tissue and FLS of model mice. And using TNF-α of 10ng/ml to stimulate FLS, and WB was taken to analysis the change of NLRC5 and IL-6, IL-1β expression level. Next, we transfected NLRC5-siRNA into the FLS with Liposome LipofectamineTM 2000, to detect the effect of TNF-α in FLS IL-6, IL-1β, p-IκBα and p-p65 expression. And finally we used the WB method to observe the expression of NLRC5 and cytokine after we use PDTC(8 μM) to stimulate the FLS for 2h.In this study, we indicated the highly expressed NLRC5 in synovium of mice RA model, which could also induce by TNF-α in FLSs. We shown that NLRC5 regulates induced IL-1β and IL-6 expression through NF-kB pathway in FLSs. Therefore, we demonstrated TNF-α induces NLRC5 expression partly associates with NF-kB pathway as well.