八肽胆囊收缩素对TNF-α诱导的大鼠滑膜细胞株RSC-364 IL-6的作用及其可能的分子机制(英文)  

Effects of cholecystokinin octapeptide on TNF-α-induced IL-6 expression and its possible molecular mechanism in rat synovial cell strain RSC-364

作　　者：赵占胜[1] 金玉怀[2] 丛斌[3] 李淑瑾[3] 徐锦荣[1] 姚玉霞[3] 凌亦凌[1] 

机构地区：[1]河北医科大学病理生理教研室,河北石家庄050017 [2]河北医科大学病原生物学教研室,河北石家庄050017 [3]河北医科大学法医学教研室,河北石家庄050017

出　　处：《中国病理生理杂志》2007年第7期1396-1401,共6页Chinese Journal of Pathophysiology

基　　金：Supported by the National Nature Science Foundation of China(No 30470679)

摘　　要：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 -364 IL-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(10-8-10-6mol/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30min,用Western blot-ting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(10-8-10-6mol/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。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.

关 键 词：关节炎 类风湿 胆囊收缩素 滑膜细胞 白细胞介素6 NF—κB 

分 类 号：R363[医药卫生—病理学]