摘要
目的研究胡椒碱(piperine,PIP)对脂多糖(LPS)活化的人结肠腺癌细胞SW480表达炎症因子的调节效应及可能的作用机制。方法采用WST-1法检测PIP对SW480细胞增殖的作用,利用流式微球捕获蛋白定量技术(cytometric beads array,CBA)检测炎症因子的蛋白表达水平,以实时定量PCR(qRT-PCR)检测炎症因子mRNA的表达水平,免疫印迹法检测p38MAPK信号通路和JNK信号通路的活化水平。结果 PIP处理可剂量依赖性地抑制SW480细胞的增殖,但PIP对细胞的毒性较小;CBA检测结果显示PIP以剂量依赖性方式抑制LPS诱导的SW480细胞中IL-8的分泌;实时定量RT-PCR检测显示,LPS+PIP处理组与LPS刺激组相比,IL-8的mRNA表达水平明显下降;免疫印迹分析表明,PIP可抑制LPS诱导的p38和JNK MAPK信号通路的活化水平。结论 PIP能够抑制LPS激活的人结肠腺癌细胞SW480中IL-8的分泌从而发挥抗炎作用,其机制可能与抑制p38和JNK MAPK信号通路有关。
This study aimed to explore the effect of piperine on the expression of inflammatory cytokines in lipopolysaccharide（LPS）-induced SW480 cells and the underlying mechanism. The effect of piperine on cell proliferation was measured by WST-1 assay; the expression of inflammatory cytokines was evaluated by cytometric beads array; and the mRNA levels of inflammatory cytokines were detected by quantitative RT-PCR. The activation of MAPK pathways（p38 and JNK） were determined by Western blotting. Our results showed that piperine not only inhibited the proliferation of SW480 cells but also significantly suppressed the expression of IL-8 protein in LPSstimulated SW480 cells in a dose-dependent manner. Moreover, piperine pretreatment could significantly depressed IL-8 mRNA expression in the LPS-induced cells, as compared to LPS treatment alone. Furthermore, MAPK pathways（p38 and JNK） that could be activated by LPS were down-regulated when the cells were pretreated with piperine. These results demonstrated that piperine could depress the expression of IL-8 in LPS-stimulated SW480 cells, both at protein and mRNA levels, and down-regulated LPS-activated MAPK pathways（p38 and JNK） which were closely related to IL-8 expression, suggesting that piperine exerted its anti-inflammatory effect by suppressing inflammation-related MAPK pathways and therefore the production of IL-8.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2014年第11期957-961,共5页
Immunological Journal
