CAJ | 학술논문

目的:观察硫酸化八肽胆囊收缩素(sCCK-8)对TNF-α诱导大鼠滑膜细胞株RSC-364IL-6mRNA表达及核因子NF-κB的影响及其可能的受体机制.方法:大鼠滑膜细胞株RSC-364经TNF-α(10μg/L)、sCCK-8(10-8-10-6 mol/L)、CCK受体拮抗剂丙谷胺(2 mg/L)及溶剂单独或联合孵育3 h,用RT-PCR检测细胞IL-6、CCK-AR及CCK-BR mRNA的表达,孵育1 h,用电泳迁移率检测NF-κB活性,孵育30 min,用Western blotting检测胞浆IκB蛋白表达.结果:RSC-364细胞固有表达CCK-A/B受体,sCCK-8(10-8-10-6 mol/L)使IL-6、CCK-AR和CCK-BR mRNA表达进一步增高,明显增加TNF-α诱导的NF-κB活性,降低胞浆中IκB蛋白水平,并可被丙谷胺所拮抗.结论:sCCK-8通过NF-κB途径上调TNF-α诱导的大鼠滑膜细胞IL-6 mRNA表达,此作用可能通过滑膜细胞上的CCK受体实现,提示CCK-8在类风湿性关节炎(RA)发病过程中可能具有调控作用.
목적:관찰류산화팔태담낭수축소(sCCK-8)대TNF-α유도대서활막세포주RSC-364IL-6mRNA표체급핵인자NF-κB적영향급기가능적수체궤제.방법:대서활막세포주RSC-364경TNF-α(10μg/L)、sCCK-8(10-8-10-6 mol/L)、CCK수체길항제병곡알(2 mg/L)급용제단독혹연합부육3 h,용RT-PCR검측세포IL-6、CCK-AR급CCK-BR mRNA적표체,부육1 h,용전영천이솔검측NF-κB활성,부육30 min,용Western blotting검측포장IκB단백표체.결과:RSC-364세포고유표체CCK-A/B수체,sCCK-8(10-8-10-6 mol/L)사IL-6、CCK-AR화CCK-BR mRNA표체진일보증고,명현증가TNF-α유도적NF-κB활성,강저포장중IκB단백수평,병가피병곡알소길항.결론:sCCK-8통과NF-κB도경상조TNF-α유도적대서활막세포IL-6 mRNA표체,차작용가능통과활막세포상적CCK수체실현,제시CCK-8재류풍습성관절염(RA)발병과정중가능구유조공작용.
AIM: To investigate the effect of sulfated cholecystokinin octapeptide (CCK -8 ) on TNF -α induced IL - 6 mRNA expression, NF - κB activation in the rat fibroblast - like synovial cell strain RSC - 364 and its possible receptor mechanisms. METHODS: RSC -364 cells were stimulated with TNF - α( 10 μg/L) in the presence or absence of sCCK- 8( 10-8 - 10-6 mol/L) or/and CCK receptor antagonist proglumide(2 mg/L). IL -6 and CCK receptor A/B (CCK- AR/CCK/BR) mRNA expression were assayed by reverse transcription polymerase chain reaction (RT- PCR) at 3 h after stimulation, and nuclear factor - κB (NF - κB) binding activity was analyzed by electrophoretic mobility shift assay (EMSA) at lh after stimulation. At 30 min of stimulation the IκB protein level in cytoplasma was measured by Western blotting. RESULTS: Both CCK - AR and CCK - BR were constitutively expressed on RSC - 364. sCCK - 8, at concentrations from 10-8 mol/L to 10 -6 mol/L, significantly increased IL - 6 mRNA expression, CCK - AR and CCK - BR mRNA expression, NF - κB binding activity and IκB protein degradation. The effects of sCCK - 8 on NF - κB activity and IκB degradation level were attenuated by CCK receptor antagonist proglumide. CONCLUSION: sCCK - 8 upregulats TNF - α- induced IL - 6 mRNA expression by NF - κB pathway through its receptor on rat synoviocytes, suggesting its possible regulatory role in the pathogenesis of rheumatoid arthritis.