不同浓度腐胺对人脐静脉内皮细胞生物学行为的影响  

Biologic effects of different concentrations of putrescine on human umbilical vein endothelial cells

作　　者：陈健霞 荣新洲[1] 樊桂成[1] 李松泽[1] 张涛[1] 李庆辉[1] 

机构地区：[1]广州医科大学附属广州市第一人民医院烧伤科,510180

出　　处：《中华烧伤杂志》2015年第6期446-450,共5页Chinese Journal of Burns

摘　　要：目的探讨不同浓度腐胺对人脐静脉内皮细胞（HUVEC）增殖、迁移、凋亡的影响。方法常规培养HUVEC，取第3～5代细胞进行以下实验。（1）按随机数字表法（分组方法下同）将细胞分为500、1000、5000μg／mL腐胺组，每组3孔，分别以含相应浓度腐胺的完全培养液培养24h后，采用倒置光学显微镜观察细胞形态。（2）将细胞分为0．5、1．0、5．0、10．0、50．0、100．0、500．0、1000．0“g／mL腐胺组及对照组，每组4孔。各腐胺组细胞采用含相应浓度腐胺的完全培养液、对照组采用不含腐胺的完全培养液培养24h后，采用比色法测定细胞增殖活性，结果以吸光度值表示。（3）同实验（2）将细胞分组（每组1孔）及培养后，Transwell迁移实验测定细胞迁移能力。（4）同实验（2）将细胞分组（每组1瓶）及培养后，采用流式细胞仪测定细胞凋亡率。对数据进行单因素方差分析、Kruskal—Wallis检验、Dunnett检验。结果（1）培养24h后，500μg／mL腐胺组细胞贴壁良好，形态无明显改变；1000μg/mL腐胺组细胞贴壁减少，细胞变小、变圆；5000μg/mL腐胺组细胞未贴壁、呈碎片状。（2）0．5、1．0、5．0、10，0、50．0、100．0、500．0、1000．0μg／mL腐胺组及对照组细胞增殖活性分别为0．588±0．055、0．857±0．031、0．707±0．031、0．662±0．023、0．450±0．019、0．415±0．014、0．359±0．020、0．204±0．030、0．447±0．021，组间总体比较差异明显（χ2=6．86，P：0．009）。0．5、1．0、5．0、10．0μg/mL腐胺组细胞增殖活性较对照组升高（P〈0．05或P〈0．01），500．0、1000．0μg／mL腐胺组细胞增殖活性较对照组降低（P值均小于0．01），50．0、100．0μg／mL腐胺组细胞增殖活性与对照组相近（P值均大于0．05）。（3）各腐胺组及对照组迁移细胞数组间总体比较差异明显（F=138．662，P〈0．001）。1．0�Objective To explore the effects of different concentrations of putrescine on proliferation, migration, and apoptosis of human umbilical vein endothelial cells （HUVECs）. Methods HUVECs were routinely cultured in vitro. The 3rd to the 5tb passage of HUVECs were used in the following experiments. （ 1 ） Cells were divided into 500, 1 000, and 5 000 μg/mL putrescine groups according to the ran- dom number table （the same grouping method was used for following grouping） , with 3 wells in each group, which were respectively cultured with complete culture solution containing putrescinc in the corresponding concentration for 24 h. Morphology of cells was observed by inverted optical microscope. （2） Cells were divided into 0.5, 1.0, 5.0, 10.0, 50.0, 100.0, 500.0, 1 000.0 μg/mL putrescine groups, and control group, with 4 wells in each group. Cells in the putrescine groups were respectively cultured with complete culture solution containing putrescine in the corresponding concentration for 24 h, and cells in control group were cultured with complete culture solution with no additional putrescine for 24 h. Cell proliferation activity （denoted as absorption value） was measured by colorimetry. （3） Cells were divided （with one well in each group） and cultured as in experiment （2） , and the migration ability was detected by transwell migration assay. （4） Cells were divided （with one flask in each group） and cultured as in experiment （2） , and the cell apoptosis rate was determined by flow eytometer. Data were processed with one-way analysis of variance, Kruskal-Wallis test, and Dunnett test. Results （1） After 24-h culture, cell attachment was good in 500 μg/mL putrescine group, and no obvious change in the shape was observed; cell attachment was less in 1 000 μg/mL putreseine group and the cells were small and rounded ; ceils in 5 000 μg/mL putrescine group were in fragmentation without attachment. （2） The absorption values of cells in 0.5, 1.0, 5.0, 10.0, 50.0, 100.0, 50

关 键 词：腐胺 内皮细胞 细胞增殖 细胞迁移分析 细胞凋亡 

分 类 号：R641[医药卫生—外科学]