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首页> 外文期刊>Life sciences >EFFECTS OF LIPOPOLYSACCHARIDE ON THE EXPRESSION OF FIBRINOLYTIC FACTORS IN AN ESTABLISHED CELL LINE FROM HUMAN ENDOTHELIAL CELLS
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EFFECTS OF LIPOPOLYSACCHARIDE ON THE EXPRESSION OF FIBRINOLYTIC FACTORS IN AN ESTABLISHED CELL LINE FROM HUMAN ENDOTHELIAL CELLS

机译：脂多糖对人内皮细胞建立的细胞系中纤溶因子表达的影响

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Human endothelial cells express antithrombotic properties by producing prostacyclin, heparan sulphate and plasminogen activator (PA). Bacterial extract, such as lipopolysaccharide (LPS), damaged the blood vessels and destroyed the balance between the antithrombotic and thrombotic functions of endothelial cells. The fibrinolytic system is involved in antithrombotic functions. The TKM-33 cell line was established from human endothelial cells. In order to determine whether TKM-33 is a good fibrinolytic system endothelial cell expression model, the expression of fibrinolytic factors in TKM-33 cells treated with or without LPS was studied. The endothelial cells which had not been treated with LPS produced and secreted a large amount of urokinase-type PA (u-PA), and small amounts of tissue-type PA (t-PA) and PA inhibitor-1 (PAI-1), which were identified immunohistochemically and by electrophoretic enzymography. Diisopropylfluorophosphate-treated I-125-u-PA bound specifically to acid-treated monolayered endothelial cells with a K-d of 2.83 +/- 0.61 nM, and B-max of (0.11 +/- 0.01) x 10(6) sites/cell. u-PAR expression was detected in endothelial cells by Northern blot analysis. Thus, endothelial cells was shown to express u-PAR which binds u-PA specifically. In the binding assay, the stimulation of endothelial cells with 0.1, 1.0 and 10 mu g/ml of LPS altered the K-d values to 6.04 +/- 0.71, 7.03 +/- 1.55 and 7.38 +/- 1.03 nM, respectively. However the B-max values did not change significantly. Although LPS treatment increased u-PAR expression in endothelial cells in a dose-dependent manner, the expression of u-PA and t-PA mRNAs was not altered significantly. LPS stimulation (10 mu g/ml) increased the expression of PAI-1 mRNA, significantly. The PA activity recovered from the cell surface fraction was not affected by LPS stimulation, but the PAI-1 activity was increased. These findings suggest that the established endothelial cell line, TKM-33, possesses the characteristics of endothelial cells and they express u-PAR on their cell surface, which is occupied by intrinsic u-PA secreted from the cells, and that treatment of endothelial cells with LPS changes the cell surface characteristics and inhibited the u-PAR expression thus promoting the prothrombotic function concomitantly with increased PAI-1 activity. [References: 36]
机译：人内皮细胞通过产生前列环素，硫酸乙酰肝素和纤溶酶原激活剂（PA）来表达抗血栓形成特性。细菌提取物，例如脂多糖（LPS），破坏了血管，破坏了内皮细胞抗血栓和血栓形成功能之间的平衡。纤溶系统参与抗血栓形成功能。从人内皮细胞建立了TKM-33细胞系。为了确定TKM-33是否是良好的纤溶系统内皮细胞表达模型，研究了用或不用LPS处理的TKM-33细胞中纤溶因子的表达。未经LPS处理的内皮细胞产生并分泌了大量的尿激酶型PA（u-PA），少量的组织型PA（t-PA）和PA抑制剂1（PAI-1） ，通过免疫组织化学和电泳酶谱鉴定。二异丙基氟磷酸盐处理过的I-125-u-PA特异性结合至酸处理过的单层内皮细胞​​，Kd为2.83 +/- 0.61 nM，B-max为（0.11 +/- 0.01）x 10（6）个位点/细胞。通过Northern印迹分析在内皮细胞中检测到u-PAR表达。因此，显示内皮细胞表达特异性结合u-PA的u-PAR。在结合测定中，用0.1、1.0和10μg/ ml LPS刺激内皮细胞将K-d值分别改变为6.04 +/- 0.71、7.03 +/- 1.55和7.38 +/- 1.03nM。但是，B-max值没有明显变化。尽管LPS处理以剂量依赖的方式增加了内皮细胞中u-PAR的表达，但是u-PA和t-PA mRNA的表达并没有明显改变。 LPS刺激（10μg / ml）显着增加PAI-1 mRNA的表达。从细胞表面部分回收的PA活性不受LPS刺激的影响，但PAI-1活性增加。这些发现表明，已建立的内皮细胞系TKM-33具有内皮细胞的特征，并且在其细胞表面表达u-PAR，该u-PAR被细胞分泌的固有u-PA占据，并且对内皮细胞的处理用LPS改变细胞表面特征并抑制u-PAR表达，从而与PAI-1活性增加同时促进血栓前功能。 [参考：36]

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