中文摘要：

目的：探讨氧化高密度脂蛋白（oxHDL）对人脐静脉内皮细胞株ECV304细胞表达组织因子（TF）的影响及其机制。方法：实验分为阴性对照组、阳性对照组（加组胺）、HDL、oxHDL4组，ECV304细胞经不同浓度oxHDL、HDL（10—120mg／L）和组胺（1×10^-9-1×10^-4mol／L）处理不同时间（2—8h）后，采用实时定量PCR（RQ—PCR）和Western bloting方法检测TF表达。同时，分别应用SP600125[C—Jun氨基端激酶（JNK）特异性抑制剂]、SB203580[038丝裂素活化蛋白激酶（p38MAPK）特异性抑制剂]、PD98059[细胞外信号调节激酶（ERKI／2）特异性抑制剂]，观察其对oxHDL和组胺诱导的ECV304细胞表达TF的影响。结果：正常ECV304细胞未检测到TF，经组胺处理1h后TFmRNA表达明显增加，在l×10^-5mol／L时最高，约为1×10^-9mol／L时的4．8倍。oxHDL以时间和浓度依赖性方式诱导ECV304细胞TFmRNA表达，在oxHDL20mg／L时最高，为10mg／L时的1．8倍，在第6h时TFmRNA表达水平为第2h的5．3倍（P〈0．05），并在20mg／L时显著刺激ECV304细胞TF蛋白表达，为40mg／L时的1．5倍，而HDL没有此效应。MAPKs（JNK、p38MAPK、ERK1／2）抑制剂可部分解除oxHDL诱导ECV304细胞TF表达的效应，TF分别下调表达至原来的19．0％、5．0％和13．0％（P〈0．05）。结论：oxHDL能以浓度和时间依赖性方式诱导ECV304细胞产生TF，其机制可能通过JNK、p38MAPK和ERK1／2所介导。

英文摘要：

AIM： To investigate the expression of tissue factor （TF） induced by oxidized high density lipoprotein （oxHDL） in human umbilical vein cell line, ECV304, and the related mechanisms. METHODS： Four main groups were designed： the negative, the positive （ECV304 with histamine）, the HDL group and the oxHDL group. Quantitative real - time polymerase chain reaction （ RQ - PCR） and Western blotting were used to detect the expression level of TF. The specific inhibitors of MAPKs, SP600125 （c -jun terminal NH2 kinase, JNK）, SB203580 （p38 MAP kinase, p38 MAPK）, PD98059 （extracellular signal -regulated kinase, ERK1/2） were used to investigate the underlying mechanisms. RESULTS： The TF expression in normal ECV304 cell line was not detected. Histamine administration resulted in a significant expression of TF in ECV304 cell line, with strongest effect after 1 h co - incubation at concentration of 1 ×10^-5 mol/L histamine （ about 4.8 - fold higher expression of TF compared with that of 1×10^-9 mol/L histamine）. Expression level of TF was detected after stimulated with oxHDL in dose - and time - dependent manners. The highest expression of TF mRNA was found at 20 mg/L oxHDL and 6 h co - incubation, with 1.8 - fold and 5. 3 - fold increase in TF expres- sion, respectively, compared with that at 10 mg/L oxHDL and 2 h co - incubation. 20 mg/L oxHDL also caused an apparent augmentation of TF protein expression, about 1.5 - fold higher compared with that stimulated by 40 mg/L oxHDL. HDL co - incubation did not cause a detectable expression of TF protein. The mRNA levels of TF in ECV304 cell line induced by oxHDL were decreased by 95.0%, 81.0%, 87.0%, respectively （all P 〈0. 05） after application of inhibitors against p38MAPK, JNK and ERK1/2. CONCLUSION： The results suggest that oxHDL stimulates TF expression in ECV304 cell line in both dose - and time - dependent manners, in which MAPKs signal transduction may play an important role.