雷公藤红素抑制血管内皮细胞增殖的AFM研究

AFM Study of the Celastrol-induced Inhibition on Vein Endothelial Cells

目的:通过抑制人脐带血管内皮细胞(HUVECs)的增殖,探讨内皮细胞对肿瘤的生长和转移的影响。方法:本实验结合原子力显微镜(AFM)、激光共聚焦显微镜(LSCM)、CCK-8和流式细胞术定性定量研究雷公藤红素(Celastrol)抑制HUVECs体外增殖。结果:0.5～2μg/ml Celastrol作用细胞24h后,CCK-8表明,细胞存活率从86.4±2.8%下降到23.4±4.8%,流式细胞术对细胞凋亡进行检测,细胞的凋亡率从11.7%上升到81.8%。AFM探测表明,随着Celastrol浓度的增大,细胞形变程度增大,细胞由长梭形逐渐变成椭圆形状,细胞表面的平均粗糙度逐渐增大。而LSCM分析细胞的骨架和伪足发现,细胞的骨架遭到破坏,伪足也逐渐消失。结论:雷公藤红素能显著抑制HUVECs的增殖。

Objective：To investigate the role of vein endothelial cells in tumor growth and metastasis,human umbilical vein endothelial cells（HUVECs） proliferation was inhibited by Celastrol.Method：Inhibition of Celastrol on HUVECs in vitro was detected by atomic force microscope（AFM）,laser scanning confocal microscope（LSCM）,CCK-8,and flow cytometer.Result：Cells were incubated with 0.5-2μg/ml Celastrol after 24h,through CCK-8 and flow cytometer assays we find that the cell viability decreased from 86.4±2.8% to 23.4± 4.8% and the apoptosis rate increased from 11.7% to 81.8% with the increase of Celastrol concentrations.The AFM results indicated that Celastrol changed the cellular morphology to various degrees and increased the surface roughness of HUVECs gradually with increasing of drug concentration.The LSCM results revealed that cytoskeleton and pseudopodium were destroyed by celastrol in a dose-dependant manner.Conclusion：HUVECs proliferation was inhibited significantly by celastrol.