基于肝窦内皮细胞功能评价隐丹参酮抑制血管新生的作用  

Evaluation of cryptotanshinone inhibition of angiogenesis in human hepatic sinusoidal endothelial cells

作　　者：黄恺 赵志敏 刘洪亮[2] 孙鑫[2] 吕靖[2] 陶艳艳[2] 刘成海[1,2,3] 

机构地区：[1]上海市中医临床重点实验室,上海201203 [2]上海中医药大学附属曙光医院肝病研究所,上海201203 [3]上海高校中医内科学E-研究院,上海201203

出　　处：《药学学报》2016年第8期1257-1262,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81173405;81473404;81473479)

摘　　要：观察隐丹参酮调控人肝窦内皮细胞(human hepatic sinusoidal endothelial cells,HHSEC)功能抑制血管新生的作用。本文以CCK8法测定隐丹参酮对HHSEC毒性;以内皮细胞生长因子(endothelial cell growth supplements,ECGS)诱导HHSEC增殖,以索拉非尼为阳性对照药,CCK8法、5-乙炔基-2'-脱氧尿苷(5-ethynyl-2'-deoxyuridine,EdU)检测细胞增殖;免疫荧光法观察胞内血管性血友病因子(von willebrand factor,v WF)的表达;荧光探针法测定胞内一氧化氮(nitric oxide,NO)水平;管腔形成实验、斑马鱼血管新生实验评价其抑制血管新生的作用。与空白组比较,ECGS显著诱导HHSEC增殖,升高vWF表达和NO水平,10μmol·L^(-1)隐丹参酮能够显著抑制HHSEC的增殖,降低vWF表达和胞内NO水平;对管腔形成及转基因斑马鱼血管新生具有显著抑制作用。结果提示,隐丹参酮具有调节内皮细胞功能抑制血管新生的活性。To investigate the effects of cryptotanshinone（an active ingredient of Salvia Miltiorrhiza） inhibition of angiogenesis, the toxicity of cryptotanshinone was assayed in human hepatic sinusoidal endothelial cells（HHSEC） by CCK8 method. Max dose without toxicity is 10 μmol·L^（-1）. The proliferation of HHSEC were induced by the endothelial cell growth supplement（ECGS）, with 2.5 μmol·L^（-1） sorafenib as the positive control. Cell proliferation was analyzed by Ed U assay. Cell viability was analyzed by CCK8 method. The expression of v WF was analyzed by immunofluorescence method. Fluorescence probe method was used to detect the intracellular nitric oxide（NO） levels. Tube formation of HHSEC and transgenic zebrafish were also observed to evaluate the effects of cryptotanshinone against angiogenesis. Compared with normal control, there is a proliferation of HHSEC induced by ECGS. The expression of v WF and the NO levels increased significantly. Cryptotanshinone inhibited the proliferation, down regulated the expression of v WF and the NO levels. Further, cryptotanshinone inhibited the tube formation of HHSEC and reduced the number of functional vessels in transgenic zebrafish. The results suggest that cryptotanshinone could inhibit angiogenesis by regulating the HHSEC cell function.

关 键 词：隐丹参酮 肝窦内皮细胞 管腔形成 斑马鱼 血管生成 肝纤维化 

分 类 号：R285[医药卫生—中药学]