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目的:探讨蛇床子素(osthole)对内毒素(lipopolysaccharide,LPS)诱导的肠上皮细胞Caco2中炎症因子表达的影响及机制。方法:培养Caco2细胞,用LPS诱导炎症反应。在LPS刺激前给予细胞不同浓度的蛇床子素处理,通过real-time PCR检测白介素(interleukin,IL)-1β、IL-6和肿瘤坏死因子(tumor necrosis factor,TNF)-α的表达情况。分别用PKA抑制剂H89和KT5720处理细胞,观察cAMP/PKA信号通路对蛇床子素效应的影响;用Western blot检测细胞中p38、Erk和JNK的磷酸化水平。结果:LPS刺激可以显著增加Caco2细胞中炎症因子IL-1β、IL-6和TNF-α的表达。蛇床子素预处理对LPS诱导的炎症反应有明显抑制作用,PKA抑制剂H89和KT5720不能逆转蛇床子素的抑制作用。LPS刺激后,Caco2细胞中p38、Erk和JNK的磷酸化水平明显增加,蛇床子素可部分抑制它们的磷酸化。结论:蛇床子素具有抑制肠上皮细胞株Caco2炎症反应的效应,该效应不依赖于cAMP/PKA,可能与抑制Erk、JNK和p38的磷酸化有关。
Objective: To investigate the effect and mechanism of osthole on the expression of inflammatory cytokines in intestinal epithelial cells Caco2 induced by lipopolysaccharide (LPS). Methods: Caco2 cells were cultured and the inflammatory response was induced by LPS. The cells were treated with different concentrations of osthole before LPS stimulation. The expression of interleukin (IL) -1β, IL-6 and tumor necrosis factor (TNF) -α were detected by real-time PCR. The cells were treated with PKA inhibitor H89 and KT5720 respectively to observe the effect of cAMP / PKA signal pathway on the osthole effect. The phosphorylation levels of p38, Erk and JNK were detected by Western blot. Results: LPS stimulation could significantly increase the expression of IL-1β, IL-6 and TNF-α in Caco2 cells. Ostikornin pretreatment significantly inhibited the inflammatory response induced by LPS, and PKA inhibitor H89 and KT5720 could not reverse the inhibitory effect of osthole. After LPS stimulation, phosphorylation levels of p38, Erk and JNK were significantly increased in Caco2 cells, and osthole partially inhibited their phosphorylation. CONCLUSION: Osthole can inhibit the inflammatory response of intestinal epithelial cell line Caco2, which is independent of cAMP / PKA and may be related to the inhibition of phosphorylation of Erk, JNK and p38.