Fujian Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350108, China.
Chin J Integr Med. 2011 Nov;17(11):840-6. doi: 10.1007/s11655-011-0895-8. Epub 2011 Nov 6.
To evaluate the angiogenic effect of the Xiongshao capsule (XSC) in human umbilical vein endothelial cells (HUVEC), and to investigate the possible molecular mechanisms mediating its biological effect.
Serum pharmacology was applied in this study, in which different doses of XSC were administrated to rats orally and then XSC-containing serum (XSC-S) was collected for the following in vitro experiments. The viability of HUVEC was determined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cell density was observed via phase-contrast microscopy. Fluorescence-activated cell sorting analysis with propidium iodide staining was performed to determine cell cycle phase. Cell migration was determined by wound-healing method. Capillary tube formation by HUVEC was examined using ECMatrix gel-based assay. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression levels were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbant assay (ELISA) analyses.
XSC-S dose-dependently stimulated proliferation of HUVEC by promoting the cell cycle G1 to S progression. In addition, XSC-S treatment dramatically increased the migration and capillary tube formation of HUVEC in a dose-dependent manner. Moreover, XSC-S enhanced the expression of VEGF and bFGF at both mRNA and protein levels.
XSC can promote several features of angiogenesis in endothelial cells through up-regulating the expression of bFGF and VEGF, suggesting that XSC may be a potential novel therapeutic agent for the treatment of ischemic heart diseases.
评价熊芍胶囊（XSC）对人脐静脉内皮细胞（HUVEC）的血管生成作用，并探讨介导其生物学效应的可能分子机制。
本研究采用血清药理学方法，将不同剂量的 XSC 灌胃给药大鼠，然后收集含 XSC 的血清（XSC-S）进行后续的体外实验。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法测定 HUVEC 的活力。通过相差显微镜观察细胞密度。采用碘化丙啶染色荧光激活细胞分选分析测定细胞周期相。采用划痕愈合法测定细胞迁移。采用 ECMatrix 凝胶基测定法检测 HUVEC 的毛细血管形成。采用逆转录聚合酶链反应（RT-PCR）和酶联免疫吸附试验（ELISA）分析测定血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）的表达水平。
XSC-S 呈剂量依赖性地刺激 HUVEC 增殖，促进细胞周期 G1 向 S 期进展。此外，XSC-S 处理以剂量依赖性方式显著增加 HUVEC 的迁移和毛细血管形成。而且，XSC-S 增强了 VEGF 和 bFGF 的表达，在 mRNA 和蛋白水平均如此。
XSC 通过上调 bFGF 和 VEGF 的表达促进内皮细胞的几种血管生成特征，表明 XSC 可能是治疗缺血性心脏病的一种有潜力的新型治疗剂。
