小鼠endostatin抑制血管内皮细胞增殖的作用机制研究

目的探讨小鼠endostatin抑制血管内皮细胞增殖的作用机制.方法在含重组小鼠 endostatin 蛋白(150 nmol/L)和小牛血清(100 ml/L)的DMEM培养基中培养人脐静脉内皮细胞ECV304, 72 h后,采用透射电镜、流式细胞仪进行细胞周期分析和细胞核DNA的1.5%琼脂糖凝胶电泳,检测重组小鼠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ECV304细胞核染色质浓缩、边集、核碎裂及胞浆浓缩等,呈典型的凋亡细胞形态学表现.流式细胞仪细胞周期分析显示,在G1期峰前存在1个凋亡峰.1.5%琼脂糖凝胶电泳显示,细胞核DNA呈梯状.对照组ECV304细胞表现正常.结论重组小鼠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.

Study on mechanism of mouse endostatin in inhibiting proliferation of vascular endothelial cells

Objective To discuss the mechanism of inhibition effects of mouse endostatin against the proliferation of vascular endothelial cells.Methods Human umbilical vein endothelial cells ECV304 were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) and recombinant endostatin protein(150 nmol/L) for 72 h. The apoptosis of ECV304 cells was detected by transmission electron microscope, flow cytometry and 1.5% agarose gel electrophoresis of nuclear DNA.Results Under the transmission electron microscope ECV304 cells treated with endostatin showed condensation and margination of nuclear chromatin, fragmentation of nuclear and condensation of cytoplasm, which were typical manifestations of apoptosis. Flow cytometry showed a visible apoptotic peak before G1 peak. The typical DNA ladder was found in 1.5% agarose gel electrophoresis of the nuclear DNA. ECV304 cells in the control group did not show these apoptotic characteristics.Conclusion The recombinant endostatin protein can induce the apoptosis of the vascular endothelial cells, which may be one of the reasons that it inhibits the proliferation of vascular endothelial cells.