论文部分内容阅读
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。
Objective To investigate the regulatory effect of piperine (PIP) on the expression of inflammatory cytokines and its possible mechanism in human colon adenocarcionoma cell line SW480 activated by lipopolysaccharide (LPS). Methods WST-1 method was used to detect the effect of PIP on the proliferation of SW480 cells. The expression of inflammatory cytokines was detected by flow cytometry beads array (CBA). The expression of inflammatory cytokines was detected by real-time quantitative PCR (qRT-PCR) mRNA expression levels, immunoblotting detection p38MAPK signal pathway and JNK signaling pathway activation level. Results PIP treatment inhibited the proliferation of SW480 cells in a dose-dependent manner, but the cytotoxicity of PIP was less. CBA assay showed that PIP inhibited LPS-induced IL-8 secretion in SW480 cells in a dose-dependent manner. PCR assay showed that IL-8 mRNA expression was significantly decreased in LPS + PIP group compared with LPS stimulation group. Western blot analysis showed that PIP inhibited LPS-induced activation of p38 and JNK MAPK signaling pathway. Conclusion PIP can inhibit the secretion of IL-8 in LPS-activated human colon adenocarcinoma cell line SW480 to exert anti-inflammatory effects, which may be related to the inhibition of p38 and JNK MAPK signaling pathway.