【摘要】 目的 探究神经营养因子Neuritin过表达对鸡胚尿囊膜（CAM）和人脐静脉内皮细胞（HUVECs）血管生成作用的影响，为治疗血管生成性疾病提供新方向。方法 选择新鲜黄皮种蛋30枚建立CAM模型，采用随机数字表法分为3组：阳性对照组（bFGF）、阴性对照组（NS）和实验组（Neuritin），每组10枚。阳性对照组载入2 500 U/mL的bFGF，实验组载入10μg/mL的Neuritin蛋白，阴性对照组载入NS，各组载入量均为10μL且所有CAM的培养温度、相对湿度及孵育时间相同，记录孵育72 h后各组CAM的血管分支、数目及大小。选择健康孕妇的新鲜脐带制作原代HUVECs，将其分为3组：转染重组质粒组（HUVEC-neu组）、转染空载体组（HUVEC-3.1组）和未转染组（HUVEC组）。HUVEC-neu组的原代HUVECs经重组质粒Neuritin转染，HUVEC-3.1组经空载体质粒转染，HUVEC组不给予特殊处理，3组均接受相同的传代培养方案。Western blot检测Neuritin在HUVEC-3.1组与HUVEC-neu组的蛋白表达水平。CCK-8实验、细胞划痕实验、Transwell实验和管型形成实验检测HUVEC-3.1组和HUVEC-neu组的细胞增殖、迁移和管型形成能力。结果 （1）实验组的CAM血管分支点数及微小血管数较阴性对照组明显增加（P <0.01），但两组的大、中血管数差异无统计学意义（P> 0.05）;(2)HUVEC-neu组中Neuritin在HUVECs成功过表达。（3）与HUVEC-3.1组相比，HUVEC-neu组的细胞增殖活力下降（P <0.05），但其迁移、管型形成能力明显增强（P <0.01）。结论 Neuritin过表达可促进血管生成，通过影响细胞增殖、迁移和管型形成能力参与新生血管的调控。更多还原

【Abstract】 Objective To investigate the effects of neurotrophic factor Neuritin overexpression on the angiogenic effects of chicken embryonic allantoic membrane(CAM) and human umbilical vein endothelial cells(HUVECs), and to provide a new direction for the treatment of angiogenic diseases. Methods Thirty fresh yellow-skinned breeding eggs were selected to establish a CAM model, which were divided into three groups by randomized numerical table method: positive control group(bFGF), negative control group(NS) and experimental group(Neuritin), with 10 eggs in each group. The positive control group was loaded with 2 500 U/mL of bFGF, the experimental group was loaded with 10 μg/mL of Neuritin protein, and the negative control group was loaded with NS. 10 μL loading volume was loaded into each group, and all CAMs were incubated at the same temperature, relative humidity, and time, and the vascular branching, number, and size of the CAMs in each group were recorded after 72 h of incubation. Fresh umbilical cords from healthy pregnant women were selected to produce primary HUVECs, which were divided into three groups: transfected with recombinant plasmid(HUVEC-neu group), transfected with empty vector(HUVEC-3.1 group), and untransfected(HUVEC group). Primary HUVECs in the HUVEC-neu group were transfected with the recombinant plasmid Neuritin, and those in the HUVEC-3.1group were transfected with the empty vector. HUVEC-3.1 group was transfected with the empty vector plasmid, and HUVEC group was not given any special treatment, and all three groups received the same culture regimen.Western blot was used to detect the protein expression level of Neuritin in HUVEC-3.1 and HUVEC-neu groups.CCK-8 assay, cell scratch assay, Transwell assay, and tube formation assay were used to detect protein expression level of Neuritin in HUVEC-3.1 group and HUVEC-neu group, and HUVEC-neu groups for cell proliferation, migration and tube formation. Results （1） The number of CAM vessel branch points and microvessels in the experimental group was significantly increased compared with that in the negative control group(P < 0.01), but there was no statistically significant difference in the number of large and medium-sized vessels between the two groups(P > 0.05);(2) Neuritin was successfully overexpressed in HUVECs in the HUVEC-neu group.(3) Compared with the HUVEC-3.1 group, the proliferation vigor of cells in the HUVEC-neu group was decreased(P < 0.05), but their migration and tube formation abilities were significantly enhanced(P < 0.01). Conclusion Neuritin overexpression promotes angiogenesis and participates in the regulation of neovascularization by affecting cell proliferation, migration, and tube formation ability.更多还原