Objective:To investigate the role of Notch signaling in lipopolysacharide (LPS) induced inflammation response in primary human umbilical vein endothelial cells (HUVEC).Methods: Primary human umbilical vein endothelial cells were cultured with or without lipopolysacharide.HUVEC were pretreated with DMSO vehicle control or γ-secretase inhibitor (DAPT) for 16 hours and subsequently stimulated with LPS.Quantitative real-time PCR were performed to measure the mRNA level of HES1.Results: LPS stimulation induced the expression of the canonical Notch target gene HES1.Peak expression of HES1 mRNA was observed at 0.5 hour after LPS stimulation, and amounts of mRNA increased in a dose-dependant manner.LPS stimulation did not increase the expression of HES1 when cells were pretreated with DAPT.Conclusions: The expression of the canonical Notch target gene HES1 is effectively induced by LPS, suggesting that the activation of Notch signaling pathway is involved in the inflammatory response induced by LPS.%目的:探讨脂多糖刺激能否诱导原代人脐静脉内皮细胞Notch信号通路的激活.方法:体外培养原代人脐静脉内皮细胞,加入脂多糖或溶媒孵育诱导炎性反应,以实时荧光定量PCR方法检测Notch信号通路靶基因HES1 mRNA的表达.予Notch信号通路阻断剂DAPT孵育16 h,再予脂多糖或溶媒刺激观察HES1 mRNA的表达.结果:脂多糖刺激诱导了Notch信号通路靶基因HES1的表达.HES1 mRNA的表达在LPS刺激后0.5 h达高峰,是对照组的11倍(P＜0.01).随着LPS浓度的增加,HES1 mRNA的表达峰值增加(P＜0.01).给予DAPT后,LPS刺激下HES1 mRNA的表达受抑制(P＜0.01).结论:首次在内皮细胞上证实LPS激活Notch信号通路靶基因HES1的表达,HES1的诱导表达依赖于经典的Notch信号通路,提示LPS的诱导在内皮细胞炎症中可能涉及Notch信号系统的激活.
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