知识与财富的链接
目的观察中药青蒿提取物青蒿琥酯(artesunate,AR)对脂多糖(lipololysaccharide ,LPS)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,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(0．04mg／L、0．2mg／L、1mg／L、5mg／L及20mg／L)共同孵育24h,ELISA方法检测培养上清中血管假性血友病因子(von Willebrand factor,vWF)含量,Western blot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1ug／ml LPS后,HUVECs的vWF及ICAM1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＜0．05)。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AR0．2mg／L及1mg／L时明显下调TNFα mR—NA表达,与LPS组比较差异有显著性(P＜0．05,P＜0．01)、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TNFα mRNA表达有关。
OBJECTIVE: To investigate the protective effects and mechanism of artesunate (AR) on the activation and injury of human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS). METHODS: After 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 expression of von Willebrand factor (vWF) in the conditioned media was tested by ELISA, the expression of intercellular adhesion molecule (ICAM-1) protein was determined by Western blot method and the expression of tumor necrosis factor alpha (TNFalpha) mRNA was determined by in situ hybridization. RESULTS: After being exposed to 1 microg/ml LPS, vWF and ICAM-1 expression were higher than those in the control group. AR could significantly down-regulate the increased expressions concentration-dependently, significant 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 could markedly down-regulate the TNFalpha mRNA expression, showing significant difference as compared with that in LPS group (P < 0.05, P < 0.01). CONCLUSION: AR has protective effect on LPS induced HUVECs activation and injury, which might be related with its inhibition on TNFalpha mRNA expression.
