Core Peptids Modulate Synovial Proliferation In Rheumatoid Arthritis

ObjectiveThe present study was conducted to investigate the consequences of corepiptide(CP) added into the co-culture system which has direct contact between Jurkat T cells and RAFS, with regard to the proliferation, apoptosis, and the expression of the cytokines of the RAFS.Methods1. RAFS were obtained by collagenase digestion, and were qualified by electron microscope and immunohistochemistry. Then, the growth and the proliferation conditions were observed.2. Jurkat T cells were cultured, the growth and the proliferation conditions of the Jurkat T cells were observed.3. MTT, flow cytometry, and real-time polymerase chain reaction analysis were used to analyze the proliferation, apoptosis. and the expression of inflammatory cytokines of Jurkat T cells.4. MTT, flow cytometry. and real-time polymerase chain reaction analysis were used to analyze the proliferation, apoptosis, and the expression of inflammatory cytokines of the Jurkat T cells and the RAFS in the co-culture system.Results1. RAFS obtained were qualified by electron microscope and immunohistrochemistry to be fibroblast like synovial cells, and they proliferated rapidly.2. The Jurkat T cells were suspended and round in the culture medium. They would get together if the culture time extended and they proliferated rapidly.3. The proliferation of the T cells was inhibited by the high dosage CP began at the7th day, the middle dosage began at the10th day, the low dosage began at the13th day. At the13th day, the inhibit effect was dose-dependent.4. After24h that CP(high dosage, middle dosage) were added, the apoptosis of Jurkat T cells, detected by cytometry, was significantly higher than the contrast group. There was no significant difference between the low dosage and the contrast. After48h, only the high dosage increased the apoptosis.5. The expression of IL-2mRNA, TNFmRNA of Jurkat T cells could be inhibited by the three dosage of CP. The expression of IL-23p19mRNA of Jurkat T cells only inhibited by the high dosage.6. At the4th day after CP added into the co-culture system, the high dosage inhibited the proliferation of the T cells. The middle and low dosage began the effect at the13th day. Only the high dosage could significantly inhibit the proliferation of RAFS.7. After24h and48h CP were added into the co-culture, all the three dosage increased the apoptosis of Jurkat T cells. The apoptosis of RAFS could be inducted by the high dosage at the24h and48h, by the middle dosage at the24h.8. The expression of IL-23p19mRNA and TNFmRNA of Jurkat T cells in co-culture system was inhibited by all the three dosage. Only the high dosage could decrease the expression of IL-23p19mRNA of RAFS in co-culture system. The middle dosage and the high dosage could decrease the expression of the TNFmRNA of RAFS in co-culture system.ConclusionCP can inhibit the proliferation, apoptosis, and the expression of IL-2mRNA, IL-23p19mRNA, TNF mRNA of the Jurkat T cells when cultured lonely or co-cultured with RAFS. High dosage of CP can inhibit the proliferation, apoptosis, and the expression of IL-23p19mRNA, TNF mRNA of RAFS when co-cultured with Jurkat T cells.