| Rheumatoid arthritis(RA), one of the connective tissue disease, has been a worldwide health concer.RA has sevious impact on the life quality and psychological health for patients and their families.RA is a chronic autoimmune disease which primarily targeds on synovium of joints. RA-FLS play an important role in the destruction of joint.The main pathological hallmark of RA include abnormal hyperplasia of synovial tissues and infiltration by inflammatory cells,synovial inflammation can lead to destructions of bone and cartilage continuously.The pathogenesis of RA is still undefined.Occurrence of RA is related to many cytokine,tumor necrosis factor-α is one of the important cytokine in RA.The most research on RA is concentrated on the abnormity of signal transduction pathway.The Wnt signaling pathway plays a key role in the synovial inflammation and the regulation of bone metabolism of RA,including the canonical and the most detailed β-catenin pathway.Since Wnt signaling is important in the regulation of RA-FLS,modulating Wnt signaling maybe a new target in searching drug treatment of RA.Resveratrol,one of polyphenolic components, has anti-oxidant、anti-aging、anti-inflammatory and cardiovascular protecting,which is widely exist in plants,such as giant knotweed rhizome、 peanut and grapes. In chinese traditional medicine,obvious effects can be found in the treatment of arthritis by giant knotweed rhizome.In recent years,Res wasobserved to induce the apoptosis of tumor cells.The proliferation and differentiation of RA-FLS are similar to tumor cells.Therefore, fibroblast-like synovial cells was used to investigate the effect of TNF-α on HFLS-RA,futher to analyze the relative mechanisms by which Res affecte the process of FLS through Wnt signal transduction pathways.Objective: 1. To observe the effect of TNF-α on the proliferation of HFLS-RA. 2. To investigate the effect of different concentration of Res on HFLS-RA proliferation induced by TNF-α. 3. To clarify the modulation of TNF-α on the Wnt signaling pathway in HFLS-RA. 4. To analyze the modulation of Res on the Wnt signaling pathway of HFLS-RA induced by TNF-αMethod: 1. To recover the HFLS-RA cell, subculture it to the 3-5th generation. 2. Adopting CCK8 method to observe the effect of TNF-α on the proliferation of HFLS-RA. 3. After HFLS- RA was treated with distinct concentration of Res(50umol/l、100umol/l、200umol/l、400umol/l)and then induced by TNF-α for 24 h, The proliferation of HFLS-RA were measured by CCK8. 4. After HFLS- RA was stimulated with TNF-α for 30 min, observeing the expression level of s FRP4、β-catenin and C-myc by Western blot betweenTNF-α group and the blank control group. 5. Adding invariable concentration of Res(50umol/l、100umol/l、200umol/l 400umol/l)respectively into HFLS-RA, incubate 24 h, then induced by TNF-α for 30 min. The expression changes of s FRP4、β-catenin and C-myc were examined by Western blot method.Results: 1. The proliferation of HFLS- RA induced by TNF-α is significantly increased. Invariable concentration of Res( 50umol/l 、 100umol/l 、 200umol/l 、400umol/l)can decrease the proliferation of HFLS-RA induced by TNF-α. The effect is dose-independent reduced. 2. The expression level of β-catenin and C-myc measured by Western blot is respectively increased in HFLS-RA after stimulated with TNF-α,but the expression of s FRP4 is significantly decreased. 3. The Western blot method shows that the expression of β-catenin、C-myc is decreased in HFLS-RA with different concentration of Res(50umol/l、100umol/l、200umol/l、400umol/l) group compared with TNF-α group, but the expression of s FRP4 is respectively increased.Conclusions: 1. TNF-α can increase the proliferation of HFLS-RA. 2. Invariable concentration of Res has inhibition effect on HFLS-RA proliferation, The effect is dose-independent reduced. 3. In HFLS-RA, the Wnt signaling pathway can be significantly activated by TNF-α. 4. Res can inhibit the activation of Wnt signaling pathway dose dependently. |
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