论文部分内容阅读
目的:观察重组血管基膜衍生多功能肽(rVBMDMP)抗血管新生的活性。方法:采用MTT法检测rVB-MDMP对血管内皮细胞增殖活性的影响,台盼蓝染色细胞计数法测定其对血管内皮细胞生长的影响,内皮管结构形成实验测定其对血管内皮细胞内皮管结构形成能力的影响,PI染色流式细胞分析术检测其对血管内皮细胞凋亡率的影响,吖啶橙染色荧光显微镜观察rVBMDMP作用后细胞凋亡形态学的改变,划痕法测定其对血管内皮细胞迁移能力的影响。结果:MTT法、台盼蓝染色细胞计数法显示rVBMDMP能显著抑制血管内皮细胞增殖和生长,其作用呈剂量和时间依赖性,1.0μmol/L rVBMDMP作用96 h时,血管内皮细胞的活细胞数仅为溶酶对照组的50%。内皮管结构形成实验结果表明,rVBMDMP能显著降低血管内皮细胞的内皮管状结构数目(17.67±3.055与50.33±3.055)。流式细胞术结果显示,rVBMDMP处理后细胞凋亡率呈浓度依赖性增加,其中1.0μmol/L浓度组凋亡率高达14.41%。吖啶橙染色荧光显微镜下观察细胞出现典型凋亡的形态学改变。划痕法显示rVBMDMP对细胞迁移能力无明显影响。结论:rVBMDMP具有显著的抗血管新生活性。
Objective: To observe the anti-angiogenic activity of recombinant vascular basement membrane-derived multi-functional peptide (rVBMDMP). Methods: The effect of rVB-MDMP on the proliferation of vascular endothelial cells was detected by MTT assay. The effect of rVB-MDMP on the proliferation of vascular endothelial cells was assayed by Trypan blue staining. The formation of endothelial cells was determined by endothelial formation assay The effect of rVBMDMP was observed by acridine orange staining fluorescent microscopy. The cell morphology of vascular endothelial cells Impact of Migration Capacity. Results: MTT assay and trypan blue staining showed that rVBMDMP could significantly inhibit the proliferation and growth of vascular endothelial cells in a dose-and time-dependent manner. When treated with 1.0 μmol / L rVBMDMP for 96 h, the number of viable cells Only 50% of the lysosomal control group. The results of endothelial tube formation showed that rVBMDMP could significantly reduce the number of endothelial tubular structures (17.67 ± 3.055 and 50.33 ± 3.055) in endothelial cells. The results of flow cytometry showed that the apoptosis rate of rVBMDMP-treated cells increased in a concentration-dependent manner, and the apoptosis rate was 14.41% in 1.0μmol / L group. Morphological changes of typical apoptotic cells were observed under acridine orange staining fluorescent microscope. Scratch assay showed that rVBMDMP had no significant effect on cell migration ability. Conclusion: rVBMDMP has significant antiangiogenic activity.