摘要
目的观察平阳霉素(PYM)对体外培养的人静脉畸形内皮细胞(human venous malformation endothelialcells,HVMECs)增殖活性和细胞形态的影响。方法采用组织块培养法,从人静脉畸形组织中培养人静脉畸形内皮细胞(HVMECs)。采用CCK-8(cell counting kit-8)试剂盒检测不同浓度PYM(0.01～1 000μg/mL)作用不同时间(6、12、18、24、48h)后HVMECs的生长抑制率。同时,通过倒置相差显微镜和扫描电镜观察内皮细胞形态。结果 0.01、1、10、1 000μg/mL PYM作用内皮细胞6h后,细胞活性为对照组的(91.3±0.4)%、(82.4±0.5)%、(75.2±0.1)%、(28.3±0.5)%。5、10、1 000μg/mL PYM作用内皮细胞24h后细胞增殖活性分别为对照组的(75.0±0.3)%、(58.4±0.5)%、(2.0±0.5)%(均P<0.01)。>10μg/mL PYM对内皮细胞活性抑制较严重。0.01μg/mL PYM作用于内皮细胞24h后,细胞未见明显改变,1μg/mL PYM作用细胞24h后,细胞内线粒体等细胞器肿胀较明显,10μg/mL PYM作用于内皮细胞24h后可见内皮细胞空泡性变,胞质水肿,核膜皱缩,染色质浓缩,核仁趋边。结论平阳霉素对人静脉畸形内皮细胞的增殖具有抑制作用,与剂量、时间呈正相关。在体外实验中PYM能诱导静脉畸形内皮细胞形态呈现凋亡样改变。
Objective To evaluate the effect of pingyangmycin（PYM）on the morphology and viability of human venous malformation endothelial cells（HVMECs）.Methods HVMECs were cultured from venous malformation tissue confirmed pathologically.Cell counting kit-8 was used to study cell viability of HVMECs treated with different concentrations of PYM（0.01-1 000 μg/mL）for different time intervals（6,12,18,24,48 h）.Morphologic changes of HVMECs were observed under phase contrast microscope and electron microscope.Results After exposure to 0.01,1,10,1 000 μg/mL PYM for 6 h,percentage of viable cells was（91.3±0.4）%,（82.4±0.5）%,（75.2±0.1）%,（28.3±0.5）% respectively as compared with the control.Cell viability was（75.0±0.3）%,（58.4±0.5）%,（2.0±0.5）% after exposure to 5,10,1 000 μg/mL PYM for 24 h（all P0.01）.Cell viability was greatly restrained at 10 μg/mL or higher concentration.No remarkable change was detected in HVMECs after exposure to 0.01 μg/mL PYM for 24 h as compared with the control.After exposure to 1 μg/mL PYM for 24 h,swelling of cell organs such as mitochondrion was detected.After exposure to 10 μg/mL PYM for 24 h,HVMECs showed vacuolar degeneration,cytoplasm swelling,wrinkled nuclear membrane,condensed chromatin,etc.Conclusion Cell viability of HVMECs was restrained by PYM in a time-and dose-depended manner.Apoptosis of HVMECs can be induced by PYM in vitro.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2011年第6期674-677,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
湖北省自然科学基金资助项目(No.2008CDZ026
No.2010CDB07907)
关键词
细胞培养
细胞活性
内皮细胞
平阳霉素
静脉畸形
cell culture
cell viability
endothelial cell
pingyangmycin
venous malformation
