青蒿琥酯对脂多糖致血管内皮细胞活化及损伤的保护作用

目的观察中药青蒿提取物青蒿琥酯 (artesunate ,AR)对脂多糖 (lipopolysaccharide ,LPS)诱导的人脐静脉内皮细胞 (humanumbilicalveinendothelialcells,HUVECs)活化及损伤状态下的保护作用及相关机制。方法建立人脐静脉内皮细胞培养 ,细胞生长至融合状态后分别加入LPS及不同浓度的AR(0 0 4mg/L、0 2mg/L、1mg/L、5mg/L及 2 0mg/L)共同孵育 2 4h。ELISA方法检测培养上清中血管假性血友病因子 (vonWillebrandfactor ,vWF)含量 ,Westernblot法检测细胞间黏附分子 (ICAM 1)蛋白表达 ,原位杂交方法检测肿瘤坏死因子 (TNFα)mRNA表达。结果暴露于 1μg/mlLPS后 ,HUVECs的vWF及ICAM 1表达较对照组明显增加 ,加入AR后 ,随AR浓度增加明显下调LPS升高的vWF及ICAM 1表达 ,至AR为1mg/L时 ,其vWF与ICAM 1表达与LPS组比较差异均有显著性 (P <0 0 5 )。AR抑制LPS升高的vWF及ICAM 1表达呈一定的浓度依赖方式。原位杂交显示在AR 0 2mg/L及 1mg/L时明显下调TNFαmR NA表达 ,与LPS组比较差异有显著性 (P <0 0 5 ,P <0 0 1)。结论青蒿琥酯对脂多糖诱导的HUVECs的活化及损伤有保护作用 ,可能与AR抑制TNFαmRNA表达有关。

英文摘要:

ObjectiveTo investigate the protective effects a nd mechanism of artesunate (AR) on the activation and injury of human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS). MethodsAfter HUVECs were cultured and turned to fusion manner, LPS and different concentration of AR (0.04 mg/L, 0.2 mg/L, 1 mg/L, 5 mg/L and 20 mg/L) were added respectively and co-incubated for 24 hrs. The expr ession of von Willebrand factor (vWF) in the conditioned media was tested by ELI SA, the expression of intercellular adhesion molecule (ICAM-1) protein was dete rmined by Western blot method and the expression of tumor necrosis factor α (TNFα) mRNA was determined by in situ hybridization. ResultsAfter being exposed to 1μg/ml LPS, vWF and ICA M-1 expression were higher than those in the control group. AR could significan tly down-regulate the increased expressions concentration-dependently, signifi cant difference showed as the concentration of AR reached 1 mg/L (P<0.05). In situ hybridization showed that AR in 0.2 mg/L and 1 mg/L cou ld markedly down-regulate the TNFα mRNA expression, showing significant differ ence as compared with that in LPS group (P<0.05, P <0.01). ConclusionAR has protective effect on LPS induced HUVEC s activation and injury, which might be related with its inhibition on TNFα mRNA expression.

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