| ObjectiveRheumatoid arthritis (rheumatoid arthritis, RA) synovitis is a kind of disease as the main manifestations of autoimmune diseases. Fibroblast-like synoviocytes are an important component of synovial tissues,which secrete high levels of pro-inflammatory cytokines,chemokines,the interaction of matrix protein degradation enzymes, continued to stimulate synovial cells,acting on the synovial signal transduction pathways in different parts,causing cells activates protein kinases, resulting in signal transduction of synovial abnormalities,as well as synovial cell proliferation and apoptosis imbalance.TNF-αhas a variety of biological activity,is an important physiological inflammatory mediators,mainly by the monocyte-macrophage cells to produce, through a unique on the target cells and play a role in receptor binding. TNFαis a center position in RA pathogenesis of pro-inflammatory cytokines involved in RA incidence of the development process.Can stimulate fibroblast-like synoviocytes proliferation,secretion of IL-6,granulocyte-macrophage colony-stimulating factor (GM-CSF), chemokines, and matrix metalloproteinase and prostaglandin and other effector molecules. In the RA pathogenesis play important role. Therefore,the role for the TNFα,TNFαantagonist, has clearance to market. But long-term use of TNF-αblockers easy to generate antibodies and cause infection. The neuropeptide CCK-8 was already results show that CCK-8 can inhibit collagen-induced arthritis fibroblast-like synoviocytes_proliferation. In RA,fibroblast-like synoviocytes_but have the same effect is not known. In our experiments, using certain concentration of CCK-8 acts on by TNFαin RA fibroblast-like synoviocytes. Observation of cell proliferation and apoptosis.MethodsRheumatoid arthritis fibroblast-like synoviocytes in primary culture:in sterile conditions to obtain fresh RA synovial tissues,isolated by collagenase digestion and purification of synovial fibroblasts,experimental generation of cells 3-7.Immunohistochemical identification of the cells cultured synovial cells.Cell growth inhibition assay (MTT) the effects of different concentrations of CCK-8 for over the role of TNFαafter the synovial cell proliferation.Experimental observation of apoptosis with different concentrations of CCK-8 for the role of TNFαafter synovial cell apoptosis.ResultsThe effect of different concentrations of CCK-8 on TNFαinduced proliferation of RAFLS.Different concentrations of CCK-8 Pre-hatch one hour after the RA FLS,the role of TNFαby adding after 48 hours,MTT results showed that TNF-α10ng/ml in the presence of, CCK-8 concentration of 10 -6 mol/L When its inhibition of the most significant difference compared with the control group significantly (P＜0.01),and other concentrations of CCK-8 compared to significant difference between (P＜0.01). The effect of CCK-8 on TNFαinduced RA FLS proliferation inhibition, with no significant positive dose-related effects. In the CCK-8 concentration of 10-6 mol/l when the most obvious of such inhibitory effect.The results showed that apoptosis CCK-8 (10 -6 mol/L) pre-incubation for an hour after the RA FLS to join the role of TNFαafter 18h,apoptosis test results showed that apoptosis rate was 61.62%,separate the role of CCK-8 of FLS, apoptosis The results showed that the apoptosis rate of 59.59%,a separate role of TNF-α(10ng/ml)of FLS, the apoptosis rate of 52.47%. The results showed that TNF-αpresence of CCK-8 can promote cell apoptosis.Conclusion1,CCK-8 can inhibit the proliferation of TNFαinduced RA FLS.2,In the CCK-8 concentration of 10 -6 mol/L, the inhibition The most obvious, but the concentration of CCK-8 was no significant positive correlation effects.3,TNF can induce the the proliferation of RA FLS, at the same time it can inhibit the apotosis of RA FLS.
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