脐静脉内皮细胞组织因子及其抑制物表达中银杏内酯B的干预效应  

Intervention of ginkgolides B for expression of tissue factor and tissue factor pathway inhibitor in human umbilical vein endothelial cells

作　　者：刘伟[1] 朱自强[1] 祖淑玉[1] 朱广瑾[1] 

机构地区：[1]中国医学科学院基础医学研究所中国协和医科大学基础医学院病理生理学系,北京市100005

出　　处：《中国组织工程研究与临床康复》2007年第6期1053-1056,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:观察银杏内酯B在氧化低密度脂蛋白诱导的脐静脉内皮细胞组织因子及其抑制物表达中的干预作用。方法:实验于2005-08/12在中国协和医科大学基础医学院病理生理学系实验室进行。采用Ⅰ型胶原酶消化分离脐静脉内皮细胞,进行原代培养至2～4代。实验分组如下:①正常对照组。②氧化低密度脂蛋白刺激组(60mg/L,氧化低密度脂蛋白作用12h)。③银杏内酯B+氧化低密度脂蛋白刺激组(预先给予20,40,80,100mg/L的银杏内酯B作用1h后,再给予60mg/L氧化低密度脂蛋白作用12h)。每组实验重复3次。分别采用发色底物法和RT-PCR技术检测脐静脉内皮细胞中组织因子和组织因子途径抑制物蛋白活性及mRNA水平的变化。结果:正常对照组脐静脉内皮细胞组织因子表达量很低,氧化低密度脂蛋白刺激脐静脉内皮细胞12h后,组织因子蛋白活性[(17.770±1.721),(2.352±0.240)U/mL,P<0.05]及mRNA水平(1.21±0.06,0.32±0.03,P<0.05)明显升高,银杏内酯B可呈剂量依赖性地抑制组织因子蛋白活性升高;组织因子途径抑制物的变化与组织因子相反,氧化低密度脂蛋白作用脐静脉内皮细胞12h后,组织因子途径抑制物蛋白活性及mRNA水平分别比对照组降低了33.8%和64.2%,但预先给予不同浓度的银杏内酯B,对组织因子途径抑制物的表达无显著影响。结论:银杏内酯B呈剂量依赖性抑制氧化低密度脂蛋白诱导的脐静脉内皮细胞组织因子蛋白活性和mRNA的表达,但对氧化低密度脂蛋白所致组织因子抑制物蛋白活性和mRNA水平的降低无显著影响。AIM： To observe the role of Ginkgolides B in tissue factor （TF） and tissue factor pathway inhibitor （TFPI） expression in human umbilical vein endothelial cells （HUVECs） induced by oxidized low density lipoprotein （oxLDL）. METHODS： The experiment was performed in the Department of Pathophysiology, institute of Basic Medical Science, Peking Union Medical College and Chinese Academy of Medical Sciences from August to December 2005. HUVECs were isolated by means of the collagenase Ⅰ perfusion technique, and the ceils were cultured 2 to 4 passages. （1） Normal control group; （2）xLDL group （60 mg/L for 12 hours）; （3）Ginkgolides B plus oxLDL group, which was treated with 60 mg/L oxLDL for 12 hours after given 20, 40, 80 and 100 mg/L Ginkgolids B for 1 hour. Every experiment was performed three times in each group. TF and TFPI protein activity and mRNA level in cultured cells were investigated with chromogenic assay and RT-PCR technique, respectively. RESULTS： TF expression in the control group was very low; After HUVECs were incubated with oxLDL for 12 hours, the protein activities [（17.770±1.721）, （2.352±0.240） U/mL, P 〈 0.05）] and mRNA levels （21±0.06, 0.32±0.03, P 〈 0.05） of TF were increased significantly, which were inhibited by Ginkgolides B in a dose-dependent manner. In contrast, after HUVECs were exposed to oxLDL for 12 hours, the protein activities and mRNA levels of TFPI were decreased 33.8% and 64.2%, respectively compared with the control group. However, the pretreatment of Ginkgolides B had no effect on the expression of TFPI. CONCLUSION： Ginkgolides B only inhibits the protein activities and mRNA levels of HUVECs induced by oxLDL in a dose-dependent manner, but not TFPI expression and mRNA levels.

关 键 词：银杏内酯B 氧化低密度脂蛋白 组织因子 组织因子途径抑制物 

分 类 号：R541.4[医药卫生—心血管疾病]