中文摘要：

目的：观察硫酸化八肽胆囊收缩素（cholecystokinin octapeptide，CCK-8）对肿瘤坏死因子α诱导大鼠滑膜细胞株RSC-364核因子κΒ的影响，以及CCK-A／B受体是否参与这一过程。 方法：实验于2003-02/2004—02在河北医科大学法医学教研室分子生物学实验室完成。取大鼠滑膜细胞株RSC-364经肿瘤坏死因子α（10μg／L）、sCCK-8（10^-8．10^-7．10^-6mol／L）、CCK受体拮抗剂丙谷胺及溶剂单独或联合应用孵育：①孵育3h，用反转录-聚合酶链反应技术检测细胞CCK-A受体及CCK-B受体mRNA的表达。②孵育1h，用电泳迁移率检测核因子κΒ相对活性。③孵育30min，用Western blot检测胞浆1κΒ蛋白表达的相对水平。 结果：①细胞CCK-A受体及CCK-B受体mRNA的表达：RSC-364细胞固有表达CCK-A／B受体，肿瘤坏死因子α（10μg/L）可使CCK-A受体和CCK-B受体mRNA的表达分别上调148％和173％（P〈0．01）。肿瘤坏死因子α和CCK-8（10^-8～10^-6mol／L）联合孵育细胞。CCK-A受体和CCK-B受体mRNA表达与肿瘤坏死因子α组相比分别增高47％,56％,30％和57％,13％,24％（P〈0．05，0．01）。②核因子κΒ相对活性：肿瘤坏死因子α组明显高于对照组（294．45±36．48,0,P〈0．01）；肿瘤坏死因子α＋CCK-8 10^-8,10^-7,10^-6mol／L组高于肿瘤坏死因子α组（470．69±56.76,489．37±64．95,558．90±74．15。P〈0．05，0．01）；CCK-8的作用可被丙谷胺减弱（400.79±39．06）。③1κΒ蛋白表达的相对水平：肿瘤坏死因子α组明显低于对照组（139．43±30．76，220.79±34．58，P〈0．01），肿瘤坏死因子α十CCK-8 10^-8,10^-7，10^-6mol／L组低于肿瘤坏死因子α组（95．28±8.54，84．15±8.77，63．28±16．13，P〈0．05），并可被丙谷胺所抑制（137．22±20．33，P〈0．01）。 结论：CCK-8对肿瘤坏死因子α诱导的RSC-364核因子κΒ活性具有正向调节作用，并能降低1κΒα蛋白水平，提示CCK-

英文摘要：

AIM： To investigate the effect of sulfated cholecystokinin octapeptide （sCCK-8） on, tumor necrosis factor-α （TNF-α） induced nuclear factor-κΒ （NF-κΒ） activity in rat flbroblast-like synovial cell strain RSC-364, and its receptor mechanisms. METHODS： The experiment was conducted in the Laboratory of Molecular Biology, Department of Forensic Medicine of Hebei Medical University between February 2003 and February 2004. RSC-364 cells were stimulated by TNF-α in the presence or absence of sCCK-8 （10^-8, 10^-7, 10^-6 mol/L）, CCK receptor antagonist proglumide and dissolvent： ① The expressions of CCK-A receptor （CCK-AR） and CCK-B receptor （CCK-BR） mRNA were assayed by reverse transcription polymerase chain reaction （RT-PCR） after 3-hour incubation. ② The NF-κΒ binding activity was analyzed by electrophoretic mobility shift assay （EMSA） after 1-hour incubation. ③After 30-minute incubation, the 1κΒ protein level in the cytoplasma was detected by Western blot. RESULTS：① mRNA expressions of CCK-AR and CCK-BR： Both CCK-ARand CCK.BRwereconstitutivelyexpressed by RSC-364. TNF-α（10 μg/L） could ,up-regulate the mRNA expressions of both CCK-AR and CCK-BR by 148% and 173% respectively （P 〈 0.01）. sCCK-8, at concentrations from 10^-8 mol/L to 10^-8 tool/L, significantly increased CCK-AR mRNA expression by 47%, 56%, 30%, and CCK-BR, 57%, 13% and 24%, after TNF-α exposure （P 〈 0.05, P 〈 0.01）. ② Relative activity of NF-κΒ： It was significantly higher in the TNF-α group than that in the control group （294.45±36.48,0, P〈 0.01 ）, and that in combination of TNF-α and sCCK-8 of all concentration group was remarkably higher than that in the TNF-α group （470.69±56.76,489.37±64.95,558.90±74.15,P 〈 0.05,0.01）. The effect of sCCK-8 was abrogated by a CCK receptor antagonist proglumide （400.79±39.06）. ③ Western Blot results： The 1κΒ protein level in the TNF-α group was obviously lower than that in the control group （139.4