脂多糖诱导血管内皮细胞凝血途径相关因子基因表达变化  

LPS changes expression of coagulation markers in human umbilical vein endothelial cells

作　　者：唐小龙[1] 蔡淑玉[1] 江振友[2] 王华东[3] 江丽芳[4] 

机构地区：[1]安徽理工大学医学院微生学与免疫学教研室,安徽淮南232001 [2]暨南大学医学院2微生学与免疫学教研室 [3]病理生理学教研室,广东广州510632 [4]中山大学中山医学院微生物学教研室,广东广州510080

出　　处：《中国病理生理杂志》2006年第8期1540-1544,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金863计划课题(No.30340013)

摘　　要：目的:观察脂多糖(LPS)对脐静脉血管内皮细胞(HUVECs)表达组织因子(TF)、组织因子抑制物(TFPI)和凝血酶调节蛋白(TM)的影响。方法:应用胰酶消化HUVECs并进行传代培养,用生长良好的第2、3代细胞进行实验。同时应用CCK-8测定细胞在不同浓度的LPS(1-100 mg/L)处理前后细胞活性;应用反转录聚合酶链反应(RT-PCR)法检测细胞内TM、TF和TFPImRNA水平。结果:浓度为10 mg/L的LPS对细胞活力与对照组相比没有显著差异。浓度为10 mg/L的LPS作用使HUVECs显著上调TF mRNA表达,6-24 h可以使细胞TFPI mR-NA表达下调,以后渐恢复正常表达,72 h达到正常对照组水平,同时下调TM mRNA表达。结论:LPS(10 mg/L)对HUVECs的活性不造成直接的影响,可显著上调HUVECs的TF mRNA转录,抑制TFPI mRNA的转录,而不改变TMmRNA的转录,这可能与LPS在感染过程中诱导血栓形成,血液凝固和D IC发生相关。AIM ： The aim of this study was to elucidate the effects of lipopolysaccharide （LPS） on the expression and secretion of tissue factor （TF） , tissue factor pathway inhibitor （TFPI） and thrombomodulin （TM） in endothelial cells. METHODS： Cultured human umbilical vein endothelial cells （HUVECs） were incubated with LPS at different concentrations of 1, 10 and 100 mg/L, respectively, for 24 hours. Cell viability was then determined by CCK- 8 assay, and lactate dehydrogenase （ LDH ） released into the culture medium was measured to assess cell damage. Levels of TF, TFPI and TM mRNA after LPS exposure were detected with reverse transcript -polymerase chain reaction （RT- PCR）. RESULTS： LPS at concentrations of 1 - 10 mg/L did not produce cell toxicity as shown by cell viability according to LDH determination in culture media after 6 h, 12 h, 24 h, 48 h and 72 h of incubation. LPS （ 100 mg/L） elicited a evident cytotoxic effect, as shown by a decreased cell viability and a higher amount of LDH in culture supernatants compared with the control cells. TF mRNA level was significantly increased when HUVECs exposed to LPS （ 10 mg/L） during 72 h. LPS treatment resulted in a marked decrease in the levels of TFPI mRNA till 24 h, then began to ascend to normal. At the same time, LPS decreased the levels of TM mRNA in HUVECs. CONCLUSIONS： A lower concentration of LPS （10 mg/L） did not show signs of cell toxicity, but it promoted the expression of TF mRNA, induced the decrease in TFPI mRNA expression and did not change TM mRNA expression. Changes of these gene expression may play an important role in the modulation of coagulation processes, which is related to thrombus and disseminated intravascular coagulation induced by LPS.

关 键 词：脂多糖类 凝血致活酶 血栓调节蛋白 脐静脉内皮细胞 基因表达 

分 类 号：R363[医药卫生—病理学]