八肽胆囊收缩素通过p38MAPK通路抑制LPS诱导的RAW264.7细胞IL-1β表达

摘要

Aim To explore the effects of cholecystokinin octapeptide ( CCK-8 ) on LPS-induced IL-1β expression in RAW264. 7 cells and its mechanism.Methods IL-1β mRNA and protein expression in LPS-induced RAW264. 7 cells were assayed by ELISA, RT-PCR and phosphorylated p38 MAPK expression was tested by Western blot. Results ① The expression of IL-1β mRNA and protein were induced by LPS in a time-dependent manner in RAW264. 7 cells, and reached peak at 3 h and 6 h respectively.②10-10 mol · L-1 CCK-8 had no effect on LPS-induced IL-1β expression in RAW264.7 cells, while 10 -8 · 10 -6 mol · L-1 CCK-8 concentration-dependently inhibits LPS-induced IL-1β expression. ③CCK-8 had no effect on LPS-induced phosphorylated p38 MAPK expression in RAW264.7 cells, while 1O-8· 10 -6 mol · L-1 CCK-8 inhibites LPS-induced phosphorylated p38 MAPK expression in a concentration-dependent manner. ④ SB203580, a specific inhibitor of p38 MAPK. could inhibit LPS-induced IL-1β expression in RAW264. 7 cells. SB203580 and CCK-8 co-incubation further inhibits LPS-induced IL-1β expression. Conclusion CCK-8 inhibits LPS-induced IL-1β exression in RAW264. 7 cells via inhibiting p38 MAPK phosphorylation, which might be one of the mechanisms of CCK-8 anti-inflammatory effects.%目的 研究八肽胆囊收缩素(CCK-8)对LPS诱导RAW264.7细胞 IL-1β表达的影响及相关机制.方法 用ELISA及RT-PCR法检测RAW264.7细胞 IL-1β mRNA及蛋白表达;用Western blot检测 RAW264.7细胞p38 MAPK的磷酸化水平.结果 ① LPS可时间依赖性的诱导RAW264.7细胞IL-1β mRNA及蛋白的表达,分别于刺激后3 h及6 h达到高峰;②10-10 mol·L-1 CCK-8对LPS诱导的RAW264.7细胞IL-1β表达无影响;10-8、10-6 mol·L-1 CCK-8浓度依赖性地抑制了LPS诱导的RAW264.7细胞IL-1β表达;③10-10 mol·L-1 CCK-8未影响LPS诱导的 p-p38 MAPK水平,10-8、10-6 mol·L-1 CCK-8浓度依赖性地抑制了LPS诱导的p-p38 MAPK水平;④ p38 MAPK特异性抑制剂SB203580可抑制LPS诱导的RAW264.7细胞IL-1β表达,与CCK-8共同作用后,抑制作用进一步加强.结论 CCK-8通过抑制p38 MAPK磷酸化而抑制了LPS诱导的RAW264.7细胞IL-1β表达,这可能是CCK-8发挥抗炎作用的信号转导机制之一.