八肽胆囊收缩素对肿瘤坏死因子-α诱导大鼠RSC-364细胞增殖及金属蛋白酶分泌的影响及作用机制  

Effect of cholecystokinin octapeptide on tumor necrosis factor-α-induced proliferation of RSC-364 cells and secretion of matrix metalloproteinases as well as relevant mechanism

作　　者：徐锦荣[1] 丛斌[2] 李淑瑾[2] 金玉怀[2] 赵占胜[2] 

机构地区：[1]河北医科大学第三医院免疫风湿科,河北石家庄050000 [2]河北医科大学法医系河北省法医学重点实验室,河北石家庄050051

出　　处：《中国生物制品学杂志》2017年第7期723-727,共5页Chinese Journal of Biologicals

基　　金：国家自然科学基金(30470679);河北省自然科学基金(C2005000705)

摘　　要：目的探讨八肽胆囊收缩素(cholecystokinin octapeptide,CCK-8)对肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)诱导大鼠RSC-364细胞增殖及金属蛋白酶(matrix metalloproteinases,MMPs)分泌的影响及作用机制。方法采用噻唑蓝(MTT)比色法检测TNF-α诱导RSC-364细胞的增殖情况,ELISA法测定细胞中MMPs(MMP-1、MMP-3、MMP-9)的分泌水平、环腺苷酸(cyclic adenosine monophosphate,c AMP)含量和蛋白激酶A(protein kinase A,PKA)活性。结果 TNF-α可促进RSC-364细胞增殖及MMP-1、MMP-3、MMP-9的分泌,CCK-8可抑制TNF-α的上述作用,Forskolin(一种c AMP诱导剂)与CCK-8的上述作用类似,而H89(PKA特异性抑制剂)可部分逆转CCK-8的上述作用。TNF-α可提高细胞中c AMP含量和PKA活性,CCK-8(10^(-10)、10^(-8)、10^(-6) mol/L)可进一步促进TNF-α的上述作用,且呈剂量依赖性,CR1409(CCK-A受体结抗剂)和CR2945(CCK-B受体结抗剂)可部分逆转CCK-8的上述作用,以CR2945为著。结论 CCK-8可通过活化c AMP-PKA信号通路,进而抑制TNF-α促细胞增殖和MMP-1、MMP-3、MMP-9的分泌,该过程由CCK受体介导。Objective To investigate the influence of cholecystokinin octapeptide（CCK-8） on tumor necrosis factor-α（TNF-α）-induced proliferation of RSC-364 cells and secretion of matrix metalloproteinases（MMPs） as well as the relevant mechanism. Methods The proliferation of RSC-364 cells induced by TNF-α was measured by monotetrazolium（MTT） colourmetric assay, while the secretion levels of MMP-1, MMP-3 and MMP-9, cyclic adenosine monophosphate（c AMP） content and protein kinase A（PKA） activity were determined by ELISA. Results TNF-α promoted the proliferation of RSC-364 cells and the secretions of MMP-1, MMP-3 and MMP-9. However, CCK-8 inhibited the abovementioned effects of TNF-α. Forskolin, a c AMP inducer, showed similar effect to that of CCK-8, while PKA specific inhibitor H89 partially reversed the effect of CCK-8. TNF-α increased the c AMP content and PKA activity in the cells,while CCK-8（10（-10）, 10（-8） and 10（-6） mol/L） enhanced the effect of TNF-α in a dose-dependent pattern. CCK-A receptor antagonist CR1409 and CCK-B receptor antagonist CR2945, especially CR2945, partially reversed the above-mentioned effect of CCK-8. Conclusion CCK-8 inhibited the TNF-α-induced proliferation of RSC-364 cells and secretion of MMPs by activating the c AMP-PKA signaling pathway in medication of CCK receptors.

关 键 词：八肽胆囊收缩素 肿瘤坏死因子-Α 类风湿关节炎 RSC-364细胞 金属蛋白酶 环腺苷酸 蛋白激酶A 

分 类 号：Q26[生物学—细胞生物学]