摘要
目的 探讨 angiostatin kringle (1- 3) [简称 AK(1-3) ]抑制血管内皮细胞增殖的作用机制 .方法 以加入重组人 AK(1- 3)蛋白 (30 0 nmol· L- 1 )的含 10 0 m L· L- 1小牛血清 DMEM培养基培养人脐静脉内皮细胞 ECV30 4,72 h后 ,采用透射电镜、流式细胞术细胞周期分析和核 DNA琼脂糖凝胶 (15 g· L- 1 )电泳检测重组 AK(1- 3)蛋白作用后血管内皮细胞的凋亡情况 .结果 实验组 ECV30 4细胞检测到了典型的凋亡 .透射电镜下可见细胞核染色质浓缩、边集、核碎裂及胞质浓缩等凋亡细胞典型的形态学改变 ;流式细胞术周期分析显示在 G1期峰前存在一个凋亡峰 (2 0 .6 % ) ;核琼脂糖凝胶(DNA15 g· L- 1 )电泳呈梯状 .对照组 ECV30 4细胞表现正常 .结论 重组人 AK(1- 3)蛋白诱导了血管内皮细胞的凋亡 。
AIM To investigate the mechanism that angiostatin kringle (1 3) [AK(1 3)] inhibits the growth of endothelial cell. METHODS Human umbilical vein endothelial cells ECV304 were cultured in DMEM medium containing 100 mL·L -1 FBS and recombinant AK(1 3) protein (300 nmol·L -1 ) for 72 h. Then the transmission electron mi croscopy, flow cytometry and 15 g·L -1 agarose gel electrophoresis of nucleus DNA were carried out to detect the apoptosis of the ECV304 cells under the effects of the AK(1 3) protein. RESULTS A typical apoptosis was detected in the ECV304 cells affected by AK(1 3) protein. By means of the transmission electron microscope we found the typical morphology variation of apoptosis, such as the concentration and side accumulation of the nuclear chromatin, the fragmentation of the nuclear and the concentration of the cytochylema etc. Cell cycle detected by flow cytometry showed one visible apoptosis peak ( 20.6% ) in front of the peak of G1. The typical ladder was found in the 15 g·L -1 agarose gel electrophoresis of the nucleus DNA. The ECV304 cells in control had no abnormal performance. CONCLUSION The recombinant AK(1 3) protein induces the apoptosis of the vascular endothelial cells, which is one reason why it inhibits the proliferation of the endothelial cells.
出处
《第四军医大学学报》
2000年第9期1051-1053,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金! (39970 85 4)
