粉防己碱抑制人脐静脉内皮细胞增殖的实验研究

Inhibitory effect of Tetrandrine on proliferation cultured human umbilical vein endothelial cell

目的观察粉防己碱(Tet)对体外培养人脐静脉内皮细胞(HUVEC)增殖的影响。探讨其抗新生血管形成的药物效应。方法以脐带来源的脐静脉内皮细胞行原代及传代培养,以光镜、第Ⅷ因子相关抗原的SABC免疫细胞方法进行观察及鉴别。应用MTT法测定不同浓度(10-3、10-4、10-5、10-6mol/L)Tet对HUVEC的抑制作用。用流式细胞仪检测Tet对细胞周期的影响。结果(1)Tet能抑制HUVEC增殖且具有量效依赖关系。(2)流式细胞仪显示HUVEC经105mol/LTet处理后,G1期增加了19·69%,S期下降了12·31%,G2期下降了7·38%。结论Tet能抑制内皮细胞增殖,是一种潜在的新生血管抑制剂。

Objective To investigate the inhibitory effect of Tetrandrine on the proliferation of human umbilical vein endothelial cell （HUVEC） in vitro. Methods Endothelial cells （EC） derived from umbilical cord veins were isolated primary and subcuhured, Light microscopy and streptoavidin biotin peroxidase complex （ SABC ） with specific antisera against Ⅷ factor were used to identify the cultured cells as epithelial cells, Cultured HUVEC were exposed to different concentrations of Tetrandrine （10^-3,10^-4,10^-5, 10^-6 mol/L） solution, and the influence of Tetrandrine on cell cycle was investigated by MTT assaying method. The changes of cell cycle were measured by flow cytometer after treatment with Tetrandrine （ 10^-5 mol/L ）. Results The OD value of HUVEC was 1,85±0.05 in 0 concentration group,and that of in 10^-3,10^-4,10^5,10^-6 mol/L of Tetrandrine groups was 0. 41±0.03,0, 56±0. 06,0. 89±0, 08 and 1.14±0. 11 , respectively, suggesting that various concentrations of Tetrandrine above mentioned significantly suppressed the proliferation of HUVEC in vitro in a dose-dependent tendency as compared to non-treatment group, The numbers of cells in G1 phase increased by 19. 69% , and that in S phase decreased by 12. 31% and 7. 38% in G2 phase after they were treated with 10^-5 mol/L of Tetrandrine,showing Tetrandrine inhibited the HUVEC division in G 0/G1 phase. Conclusion Inferring Tetrandrine may be a potent antiangiogenic agent because of inhibition of cellular proliferation.