CCK-8降低TNF-α诱导的大鼠RSC-364细胞增殖及p38 MAPK活性

CCK-8 decreases RSC-364 proliferation and p38 MAPK activation induced by TNF-α in rats

目的 观察硫酸化八肽胆囊收缩素（CCK-8）对TNF-α诱导大鼠成纤维样滑膜细胞株RSC-364细胞增殖及丝裂原活化蛋白激酶p38（p38 MAPK）活性的影响.方法 采用噻唑蓝（MTT）比色法检测细胞增殖;Western blot技术检测p38 MAPK活性.结果TNF-α（50 μg/L）孵育5 min,p38 MAPK磷酸化水平升高,15 min达高峰,2 h恢复基础水平;孵育15 min时,p38 MAPK磷酸化程度随其剂量（10、25、50 μg/L）的增大而增加.CCK-8（10^-10～10^-6 mol/L）剂量依赖性降低TNF-α诱导的细胞增殖及磷酸化p38 MAPK的激活,此作用可被CR1409和CR2945拮抗.SB203580（10 μmol/L）可抑制TNF-α引起的细胞增殖.结论 CCK-8通过降低p38 MAPK磷酸化水平而抑制TNF-α激活的RSC-364细胞增殖,该作用可能由CCK-A和CCK-B受体共同介导.

Objective To study the effect of cholecystokinin-octapeptide （CCK-8） on the proliferation of fibroblast-like synovial call line RSC-364 and p38 MAPK activity induced by TNF-α in rat. Methods The proliferation of RSC-364 cells was measured by monotetrasolium （MTT） colourmetric assay and the level of activation of p38 MAPK was deteced by Western blot. Results An increase in p38 MAPK phosphorylation was detected 5 min after TNF-α（50 μg/L）addition, and reached a plateau at 15 min, finally returned to the basic level at 2 h. TNF-α（10, 25, 50 μg/L） increased p38 MAPK phosphorylation in a dose dependent manner at 15 min. CCK-8 （ 10- ,0 ^-10- 10^-6 mol/L） could inhibit the proliferation and the level of phosphorylation of p38 MAPK in a dose dependent manner. Moreover the inhibitory effects were partly reversed by CCK-A receptor specific antagonist CR1409 or CCK-B receptor specific antagonist CR2945. SB203580 inhibited TNF-α-stimulated RSC-364 proliferation. Conclusion CCK-8 inhibited TNF-α-stimulated proliferation by decreasing p38 MAPK phosphorylation in RSC-364 cells, which was mediated through CCK-A receptor or CCK-B receptor.