Cultivated Orostachys japonicus extract inhibits VEGF-induced angiogenesis via regulation of VEGFR2 signaling pathway in vitro and in vivo

• Published in: Journal of ethnopharmacology 2020-06, Vol.256, p.112664-112664, Article 112664
• Main Authors: Cho, Hyun-Dong, Lee, Kwan-Woo, Won, Yeong-Seon, Kim, Jeong-Ho, Seo, Kwon-Il
• Format: Article
• Language: English
• Subjects: Angiogenesis; Angiogenesis Inducing Agents - pharmacology; Angiogenesis Inhibitors - pharmacology; Animals; Cell Cycle Checkpoints - drug effects; Cell Line; Cell Movement - drug effects; Cell Proliferation - drug effects; Collagen - drug effects; Collagen - metabolism; Crassulaceae - chemistry; Cultivated Orostachys japonicus; Drug Combinations; G1 Phase - drug effects; Human Umbilical Vein Endothelial Cells; Humans; Laminin - drug effects; Laminin - metabolism; Male; Mice; Mice, Inbred C57BL; Neovascularization, Pathologic - drug therapy; Neovascularization, Pathologic - metabolism; Plant Extracts - pharmacology; Proteoglycans - drug effects; Proteoglycans - metabolism; Rats; Rats, Sprague-Dawley; Resting Phase, Cell Cycle - drug effects; Signal Transduction - drug effects; Vascular Endothelial Growth Factor A - metabolism; Vascular endothelial growth factor receptor 2; Vascular Endothelial Growth Factor Receptor-2 - metabolism
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2020.112664

Orostachys japonicus A. Berger (O. japonicus), so-called Wa-song in Korea, a traditional food and medicine that grows on mountain rocks and roof tiles. Wa-song containing various phenolic compounds have been reported as a medicinal plant for prevention of fibrosis, cancer, inflammation, and oxidative damage. The present study was designed to examine the anti-angiogenic effects of cultivated Orostachys japonicus 70% ethanol extract (CE) in vascular endothelial growth factor (VEGF)-stimulated human umbilical vein endothelial cells (HUVECs). CE was prepared with 70% ethanol. HUVECs, rat aortic rings, and matrigel plug in mice were treated with CE (10–20 μg/mL) and VEGF (20–50 ng/mL), and the anti-angiogenic activities of CE were analyzed by SRB, wound healing, trans-well invasion, capillary-like tubule formation, rat aortas, Western blot, and matrigel plug assay. Phenolic compounds in CE were analyzed using a high-performance liquid chromatography (HPLC)-PDA system. Treatment of CE (10–20 μg/mL) markedly suppressed proliferation of HUVECs in the presence (from 136.5% to 112.2%) or absence of VEGF (from 100.0% to 92.1%). The proliferation inhibitory effect of CE was caused by G0/G1 cell cycle arrest, and the decrease of CDK-2, CDK-4, Cyclin D1 and Cyclin E1. Furthermore, CE treatment showed significant angiogenesis inhibitory effects on motility, invasion and micro-vessel formation of HUVECs, rat aortic rings and subcutaneous matrigels under VEGF-stimulation condition. In HUVECs, CE-induced anti-angiogenic effect was regulated by inhibition of the PI3K/AKT/mTOR, MAPK/p38, MAPK/ERK, FAK-Src, and VEGF-VEGFR2 signaling pathways. This study demonstrated that CE might be used as a potential natural substance, multi-targeted angiogenesis inhibitor, functional food material. [Display omitted] •CE blocked cell cycle progression in G0/G1 phase.•CE inhibited VEGF-induced endothelial cell migration, invasion, and differentiation into capillary-like structure in vitro.•CE inhibited phosphorylation of PI3K/AKT/mTOR and MAPK signaling pathways in HUVECs.•CE inhibited activation of VEGFR2.•CE suppressed VEGF-induced angiogenesis in vivo.