青蒿琥酯对脂多糖诱导人结肠上皮细胞炎症反应影响的分子机制研究

Effects of artesunate on inflammatory reaction of LPS-induced human colon epithelial cells and its molecular mechanism

目的探讨青蒿琥酯对脂多糖诱导人结肠上皮细胞炎症反应的影响,并研究青蒿琥酯对人结肠上皮细胞炎症治疗作用的可能机制。方法体外原代培养人结肠上皮细胞,并分为空白对照组,LPS诱导组,青蒿琥酯低、中、高剂量组,CCK-8法检测青蒿琥酯对人结肠细胞增殖的影响,ELISA法检测青蒿琥酯对人结肠上皮细胞分泌IL-6和TNF-α炎症因子的影响,Western blot检测青蒿琥酯对Traf6/p65信号通路的影响。结果与LPS诱导组相比,青蒿琥酯能显著改善LPS对人结肠上皮细胞增殖的抑制作用(P<0.05);与LPS诱导组相比,各剂量青蒿琥酯均显著降低了人结肠上皮细胞分泌的IL-6和TNF-α水平(P<0.05);免疫印迹法检测发现,各剂量青蒿琥酯组相对LPS诱导组细胞中Traf6和p-p65表达水平明显降低(P<0.05)。结论青蒿琥酯可抑制人结肠上皮细胞炎症反应,进而改善炎症对细胞增殖的抑制作用,其作用机制可能与下调Traf6/p65信号通路活化有关。

Objective To explore the effects of artesunate on inflammatory reaction of lipopolysaccharide（ LPS）-induced human colon epithelial（ HCE） cells,and study the potential mechanism of artesunate treatment for HCEC inflammation. Methods Primary human colon epithelial cells were cultured in vitro,and HCE cells were divided into five groups,including control group,LPS-induced group,low-dose-,medium-dose-,and high-dose-artesunate groups. The effects of artesunate on the proliferation of HCE cells were observed using CCK-8 assay. ELISA was used to investigate the influence of artesunate on the secretory levels of inflammatory cytokine（ IL-6 and TNF-α）. Western blot was used to assess the impacts of artesunate on the Traf6/p65 signaling pathway. Results Artesunate could significantly improve the inhibitory effect on LPS-induced HCE cells proliferation（ P〈0. 05）. Varying concentrations of artesunate could significantly inhibit the secretion of IL-6 and TNF-α in HCE cells,compared with LPS-induced group（ P〈0. 05）. Western blot results showed that the expression levels of Traf6 and p-p65 in all artesunate groups were lower than those of LPS-induced group（ P〈0. 05）. Conclusion Artesunate can significantly inhibit the inflammatory reaction of HCE cells and its molecular mechanism may involve the down-regulation of activation of Traf6/p65 signaling pathway.