人海绵状血管瘤动物模型的建立和治疗药物的筛选  

Establishment of an animal model of human cavernous angioma for drug screening

作　　者：张文健[1] 吴练秋[1] 顾锋[2] 叶丽亚[1] 辛育龄[3] 杨治华[4] 娄晋宁[1] 

机构地区：[1]中日友好医院临床医学研究所,北京100029 [2]中国医学科学院北京协和医院内分泌科 [3]中日友好医院血管外科 [4]中国医学科学院肿瘤研究所

出　　处：《中国医药生物技术》2007年第1期13-17,共5页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(30230150);国家重点基础研究发展计划(973计划)(2002CB513100-6)

摘　　要：目的建立人海绵状血管瘤的动物模型,并用于药物筛选。方法用荆豆凝集素包被的免疫磁珠分离和纯化人海绵状血管瘤内皮细胞(HCAEC)。将HCAEC(2.5×106个)与人肝癌细胞Bel-7402(5×105个)混合接种于裸鼠皮下,建立海绵状血管瘤的动物模型。通过绿色荧光蛋白示踪和组织学检查分析血管瘤中内皮细胞的来源和血管瘤的病理特征。另外,应用血管瘤内皮细胞和血管瘤动物模型筛选血管瘤的治疗药物。结果HCAEC与Bel-7402细胞共移植于裸鼠皮下后7～9d即可见接种局部形成血管瘤样结构。组织学检查显示血管瘤模型的组织学特征与人的海绵状血管瘤非常相似。细胞绿色荧光蛋白示踪显示绿色荧光蛋白存在于血管瘤的管壁,表明这些细胞来源于接种的HCAEC。药物筛选发现876-3,一种从中药中提取的单体,体外明显抑制HCAEC增殖,体内对血管瘤具有良好治疗作用。结论HCAEC与肝癌细胞共移植在裸鼠皮下可以形成人源化血管瘤动物模型,这种血管瘤动物模型可以用于治疗药物的筛选。Objective To establish an animal model of human cavernous angioma for drug screening. Methods Endothelial cells derived from human cavernous angioma (HCAEC) were isolated and purified by Ulex europaeus agglutinin-1(UEA-1)-conjugated magnetic beads. An animal model of human cavernous angioma was established by co-inoculating HCAEC and human liver cancer cells (Bel-7402) into immune-deficient nude mice subcutaneously. GFP-tagging and histopathological examinations were used to analyze the source of endothelial cells and pathological features in the angioma tissue. In addition, drugs for angioma were screened by HCAEC assay in vitro, and using the angioma model in vivo. Results An angioma-like structure was found on the site of co-inoculation of HCAEC (2.5×106) and Bel-7402 cells (5×105) 7-9 days after the transplantation. The histological feature of the angioma-like tissues was similar to that of a human angioma. GFP-tagging showed green fluorescence on the vascular wall of the angioma-like tissues, suggesting that the endothelial cells came from the inoculated HCAEC. Drug screening found that 876-3, an extract from a Chinese traditional medicine, was an effective drug for inhibiting the proliferation of HCAEC in vitro, and for treating the angioma in vivo. Conclusions An animal model of human angioma can be established by co-inoculating HCAEC and Bel-7402 cells into immune-deficient nude mice, and this animal model can be used in drug screening for angioma..

关 键 词：血管瘤 海绵状 内皮细胞 动物模型 药物评价 临床前 

分 类 号：R732.2[医药卫生—肿瘤]