血管内皮生长因子和微血管密度在接种肝泡状棘球蚴沙鼠体内的表达及意义  

Expression of vascular endothelial growth factor and microvessel density in gerbil model with inocu-lated hepatic alveolar echinocoecsis]

作　　者：李琪[1] 金亮[1] 蔡飞[1] 陈骞[1] 张怀[1] 张示杰[2] 

机构地区：[1]石河子大学医学院,新疆832000 [2]石河子大学医学院第一附属医院肝胆外科

出　　处：《中华肝胆外科杂志》2016年第4期268-272,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：基金项目：国家自然科学基金（81560518）

摘　　要：目的研究血管内皮生长因子（VEGF）、微血管密度（MVD）-CD34在接种肝泡状棘球蚴（HAE）沙鼠体内的表达及意义。方法将60只健康长爪沙鼠随机平均分为3组，即模型组、对照组和假手术组。模型组沙鼠采用开腹肝穿刺法，每只鼠接种泡球蚴原头节混悬液0．1ml。对照组以相同方法接种等量磷酸缓冲盐溶液（PBS），假手术组开腹肝穿后直接缝合。接种后120天，眼球取血。模型组取泡球蚴组织、泡球蚴周围肝组织和远端肝组织，剩余两组取肝组织，以苏木精．伊红（HE）染色、免疫组化、蛋白印迹及酶联免疫吸附法（Elisa）分析YEGF及CD34的表达。结果模型组肝脏中可见大小不等的囊泡组织，部分播散至腹腔。阳性VEGF呈黄色或棕黄色颗粒，主要存在于胞质，偶见于包膜。泡球蚴组织、泡球蚴周围肝组织和远端肝组织以及对照组和假手术组肝组织VEGF评分和VEGF／β-肌动蛋白比值分别为5．13±1．81和1．12±0．05、4．20±1．52和0．91±0．02、2．464-1．06和0．28±0．02、2．20±0．77和0．30±0．02、2．13±1．72和0．25±0．01。VEGF的表达随组织部位的不同呈梯度改变，在泡球蚴组织中表达最高（P〈0．05）。阳性CD34颗粒呈黄褐色，主要位于内皮细胞的细胞膜。泡球蚴组织、泡球蚴周围肝组织CD34计数和CD34／β-actin比值分别为：5．06±2．46、0．66±0．01和2．43±1．26、0．58±0．02，以泡球蚴组织中表达最高（P〈0．05），其余部位未检测到CD34阳性表达。模型组、对照组、假手术组沙鼠血清VEGF和CD34的表达分别为（106．17±14．17）pg／ml和（1．47±0．28）ng／ml、（38．13±21．36）pg／ml和（0．82±0．12）ng／ml、（42．88±17．25）pg／rnl和（0．72±0．16）ng／ml。模型组沙鼠血清VEGF和CD34含量明显高于对照组与假手术组（P〈0．05）。结论沙鼠HAE组织存在血管新生迹象，而血管新�Objective To study the expression of vascular endothelial growth factor （VEGF） and mierovessel density （ MVD, CD34） in inoculated hepatic alveolar eehinococesis gerbil model and explore the clinical significance. Methods Sixty healthy gerbils were randomly divided into model group, control group and sham operation group. Each gerbil in both model and control groups had liver vaccination with 0.1 ml immature alveolar echinococcus protoscolex suspension or 0.1 ml phosphate buffered saline by open abdomi- nal surgery, respectively, and the sham operation group only had abdominal surgical procedures. Gerbils were put to death on the 120th day post operation, extracted the eyeball and drawn the blood. The alveolar hydatid tissue, surrounding amt distal liver tissue were removed from the model group, and normal liver tis- sue was collected tYom the other two groups. The VEGF and MVD-CD34 expression was detected by immu- nohistochemistry staining, Western Blot, and enzyme-linked immunosorbent assay （Elisa）, Results Liver alveolar echinococcsis tissue was only observed in model group. The VEGF expression was observed as yellow or brown yellow granules located in the cytoplasma. The VEGF score and VEGF/β-actin ratio in thedifferent parts were 5.13 ± 1.81 and 1.12 ±0. 05, 4. 20 ±1.52 and 0. 91 ±0. 02, 2. 46 ±1.06 and 0. 28 ± 0. 02, 2. 20 ±0. 77 and 0. 30 ±0. 02, 2. 13 ± 1.72 and 0. 25 ±0. 01, respectively. The VEGF expression in different histological sites showed a gradient change, and bubble hydatid tissues had the highest VEGF ex- pression （P 〈 0. 05 ）. CD34 was expressed in alveolar hydatid tissue and surrounding liver tissue in model group and the CD34 positive cells was stained brown, while CD34 was mainly located in the cell membrane of endothelial cells. The CD34 scores and the CD34/β-aetin ratio in the different parts were 5.06 ± 2. 46 and 0. 66 ±0. 01, 2. 43 ± 1.26 and 0. 58 ±0. 02, and the other tissues did not have CD34 expression. The alveolar hydatid tissue had higher CD34 ex

关 键 词：泡状棘球蚴 肝 血管内皮生长因子 微血管密度 血管新生 沙鼠 

分 类 号：R532.32[医药卫生—内科学]