脂多糖诱导大鼠主动脉内皮细胞表达组织因子的体外研究

目的选择最佳浓度的脂多糖（LPS）诱导大鼠主动脉内皮细胞（RAECs）表达组织因子（TF）,并观察TF表达变化,找出TF表达最强的作用时间,从而建立高度表达TF的RAECs受损模型。方法将0.1、1、10和100μg/mlLPS分别作用RAECs6、12、24、48h,应用MTT比色法检测细胞活力,选择最佳浓度LPS作用RAECs不同时间,并应用免疫组织化学法和Western blotting法检测RAECs内TF蛋白水平。结果24h内,浓度为0.1和1μg/mlLPS对细胞活力影响与阴性对照组相比,差异无统计学意义（P〉0.05）。1μg/mlLPS能显著上调RAECsTF蛋白水平,4h左右TF蛋白表达最强,以后表达逐渐下降至正常水平。结论1μg/mlLPS作用RAECs4h左右TF蛋白表达最强,成功建立高度表达TF的RAECs受损模型。

Study of Expression of Tissue Factor in RAECs Induced by LPS in Vitro

Objective To observe the expression of tissue factor（TF） in rat aortic endothelial cells （RAECs） induced by optimal dose of lipopolysaceharides（LPS） for different duration and to find the action time when expression of TF reached the peak level. Methods Cultured RAECs were incubated with LPS at different concentrations of 0. 1 μg/ml, 1 μg/ml, 10 μg/ml and 100 μg/ml, respectively, for 6, 12, 24, 48 hours, cell viability was then determined by MTF, level of TF protein was detected with immunohistochemistry and Western blotting methods. Results LPS at concentrations of 0.1μg/ml and 1μg/ml did not produce cell toxicity as shown by cell viability. TF protein level was significantly increased after 1 μg/ml LPS exposure, and reached the peak at about 4h, then began to descend to normal. Conclusion 1μg/ml LPS promoted the expression of TF protein, and the peak value occured at about 4h.