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目的 探讨人endostatin抑制血管内皮细胞增殖的作用机制。方法 在含重组人endostatin蛋白和 10 %小牛血清的DMEM培养基中 ,培养人脐静脉内皮细胞ECV30 4。72h后 ,采用透射电镜、流式细胞术细胞周期分析和细胞核DNA的 1.5 %琼脂糖凝胶电泳 ,检测重组人endostatin蛋白作用后 ,血管内皮细胞的凋亡。结果 透射电镜下可见实验组ECV30 4细胞核染色质浓缩、边集、核碎裂及胞浆浓缩等 ,呈典型的凋亡细胞的形态学表现。流式细胞术细胞周期分析显示 ,在G1期峰前存在 1个凋亡峰 (2 0 .6 % )。 1.5 %琼脂糖凝胶电泳显示 ,细胞核DNA呈梯状。对照组ECV30 4细胞表现正常。结论 重组人endostatin蛋白可诱导血管内皮细胞凋亡 ,是其抑制血管内皮细胞增殖的原因之一。
Objective To investigate the mechanism of human endostatin inhibiting the proliferation of vascular endothelial cells. METHODS: Human umbilical vein endothelial cells (ECV30) were cultured in DMEM medium containing recombinant human endostatin protein and 10% fetal bovine serum for 4.72 h. The cell cycle of the cells was analyzed by flow cytometry and 1.5% agarose Gel electrophoresis was used to detect the effect of recombinant human endostatin protein on the apoptosis of vascular endothelial cells. Results The results of transmission electron microscope showed that the chromatin condensation of ECV30 4 cells in nucleus testis, nucleus fragmentation and cytoplasm concentration showed the typical morphological changes of apoptotic cells. Flow Cytometry Cell cycle analysis showed that there was a peak of apoptosis (20.6%) before G1 peak. 1.5% agarose gel electrophoresis showed nuclear DNA was ladder-shaped. The control group ECV30 4 cells showed normal. Conclusion Recombinant human endostatin protein can induce the apoptosis of vascular endothelial cells, which is one of the reasons that it inhibits the proliferation of vascular endothelial cells.