Effect of natural hirudin on angiogenesis of human microvascular endothelial cells

• Published in: Zhongguo xiu fu chong jian wai ke za zhi 2018-12, Vol.32 (12), p.1586-1591
• Main Authors: Lin, Guanyu, Lin, Bojie, Zhu, Jiangying, Pan, Xinyuan, Yin, Guoqian
• Format: Article
• Language: Chinese
• Subjects: Cell Proliferation; Cells, Cultured; Endothelial Cells - drug effects; Endothelial Cells - physiology; Hirudins - physiology; Humans; Neovascularization, Physiologic - drug effects; Vascular Endothelial Growth Factor A - metabolism
• ISSN: 1002-1892
• DOI: 10.7507/1002-1892.201806055

To explore the effect of natural hirudin on proliferation of human microvascular endothelial cells (HMVECs) and its preliminary mechanism of promoting angiogenesis. Three-dimensional culture models of HMVECs were established and observed by inverted phase contrast microscopy after 24 hours of culturing. Then, the three-dimensional culture models of HMVECs were treated with different concentrations (1, 4, and 7 ATU/mL) of the natural hirudin, respectively, and Dulbecco's modified Eagle's medium containing 10% fetal bovine serum as control. The cell proliferations of 4 groups were detected by cell counting kit 8 (CCK-8) method at 24, 48, and 72 hours; the angiogenesis of 4 groups were observed by tube formation assay at 24 hours; the expressions of vascular endothelial growth factor (VEGF) and Notch1 of HMVECs in 4 groups were observed by immunofluorescence staining at 24 hours. The observation of cells in three-dimensional culture models showed that HMVECs attached to Matrigel well, and the cells formed tube structure completely after 24 hours. The results of CCK-8 test showed that the absorbance ( ) value of 1 and 4 ATU/mL groups were higher than that of control group at each time point (