雷公藤甲素对血管内皮细胞尿激酶型纤溶酶原激活物表达的影响  

Effect of Triptolide on Expression of Urokinase-type Plasminogen Activators in Vascular Endothelial Cells

作　　者：丁怡[1] 张杰[2] 赵庆亮[2] 侯立军[2] 王秋林[1] 王树人[1] 

机构地区：[1]四川大学华西基础与法医学院病理生理教研室,四川成都610041 [2]潍坊医学院眼科,山东潍坊261042

出　　处：《癌症》2005年第7期801-805,共5页Chinese Journal of Cancer

基　　金：山东省教育厅重点资助课题(No.J01K08)~~

摘　　要：背景与目的已有研究发现雷公藤具有抗肿瘤的作用,本研究旨在观察雷公藤甲素(triptolide,T10)对血管内皮细胞增殖和尿激酶型纤溶酶原激活物(urokinase-typeplasminogenactivator,u-PA)表达的影响,探讨T10抑制血管生成的机制。方法体外培养人脐静脉内皮细胞,传至第三代,加入不同浓度的T10(0、5、10、20、30μg/L)、地塞米松(300mg/L)后培养不同时间,计数细胞以了解细胞的增殖情况;培养48h,采用[3H]-TDR掺入法检测DNA的合成情况;利用鸡胚尿囊膜在体观察T10对血管生成的影响;免疫组织化学染色检测内皮细胞u-PA的蛋白表达;荧光定量RT-PCR检测u-PAmRNA的表达。结果T10可抑制内皮细胞的增殖,并且呈明显的剂量依赖性,5μg/LT10的抑制率达28.93%;可明显抑制鸡胚尿囊膜的血管生成(P<0.05);减少内皮细胞u-PA的蛋白表达,并呈剂量依赖趋势,5μg/LT10使内皮细胞u-PA表达降低41.05%;荧光定量RT-PCR结果显示,T10可下调u-PAmRNA的表达。结论雷公藤甲素可有效抑制血管内皮细胞的增殖,减少内皮细胞u-PA的表达,这可能是其抑制血管生成的主要机制之一。BACKGROUND & OBJECTIVE: It has been showed that triptolide (T10) has antitumor activities. This study was to observe the inhibitory effects of T10 on proliferation of vascular endothelial cells and expression of urokinase-type plasminogen activator (u-PA), and to elucidate the antiangiogenic mechanism of T10. METHODS: Human umbilical vein endothelial cells were cultured in vitro for 3 generations, and treated with T10 (0, 5, 10, 20, and 30 μg/L), or dexamethasone (300 mg/L) as control. Cell count and 3H-TDR incorporation were used to observe cell proliferation. The chick embryo chorioallantoic membrane (CAM) test was carried out to observe the effect of T10 on angiogenesis. Immunohistochemistry and real-time quantitive reverse transcription-polymerase chain reaction (RT-PCR) were used to detect protein and mRNA levels of u-PA. RESULTS:T10 inhibited the proliferation of umbilical vein endothelial cells in a dose-dependent manner; inhibitory rate of endothelial cells was 28.93% after treatment of 5 μg/L of T10. T10 significantly reduced angiogenesis in CAM, even at the concentration of 5 μg/L (P<0.05). T10 decreased protein and mRNA levels of u-PA in endothelial cells in a dose-dependent manner; the protein level of u-PA was reduced by 41.05% after treatment of 5 μg/L of T10. CONCLUSION: T10 could inhibit the proliferation of human umbilical vein endothelial cells and expression of u-PA, which may contribute to its antiangiogenic activity.

关 键 词：雷公藤甲素/药理学 血管内皮细胞 尿激酶型纤 溶酶原激活物 增殖 血管生成 

分 类 号：R285[医药卫生—中药学]