The antiangiogenic activity of Kushecarpin D, a novel flavonoid isolated from Sophora flavescens Ait

• Published in: Life sciences (1973) 2013-11, Vol.93 (21), p.791-797
• Main Authors: Pu, Li-Ping, Chen, He-Ping, Cao, Mei-Ai, Zhang, Xiu-Li, Gao, Qing-Xiang, Yuan, Cheng-Shan, Wang, Chun-Ming
• Format: Article
• Language: English
• Subjects: Angiogenesis Inhibitors - isolation & purification; Angiogenesis Inhibitors - pharmacology; Antiangiogenic; apoptosis; Apoptosis - drug effects; Benzofurans - isolation & purification; Benzofurans - pharmacology; Benzopyrans - isolation & purification; Benzopyrans - pharmacology; catalase; Catalase - metabolism; cell adhesion; Cell Adhesion - drug effects; cell cycle; Cell Cycle - drug effects; Cell cycle arrest; cell movement; Cell Movement - drug effects; cell proliferation; Cell Proliferation - drug effects; colorimetry; Drugs, Chinese Herbal - pharmacology; endothelial cells; Flavonoid; flavonoids; Flavonoids - pharmacology; flow cytometry; fluorescence; herbal medicines; Human Umbilical Vein Endothelial Cells; Humans; Kushecarpin D; mechanism of action; medicinal plants; molybdates; Plant Roots - chemistry; Reactive oxygen species; Reactive Oxygen Species - metabolism; Sophora - chemistry; Sophora flavescens; Sophora flavescens Ait
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2013.09.025

Kushecarpin D (KD) is a novel flavonoid isolated from the traditional Chinese herbal medicine Kushen (the dried root of Sophora flavescens Ait). As part of our continuous effort to explore Chinese traditional medicinal herbs and to identify novel natural anticancer products, the antiangiogenic properties of KD were examined in vitro using a human umbilical vein endothelial cell line (ECV304). The SRB and Trypan Blue exclusion assays were used to evaluate the effect of KD on cell proliferation. The antiangiogenic activities of KD were evaluated through studies of cell migration, cell adhesion, and tube formation. DCFH-DA and DHE fluorescent assays were used to detect the reactive oxygen species (ROS) levels. Catalase activity was detected using the colorimetric ammonium molybdate method. Cell cycle and apoptosis were measured using flow cytometry and the Hoechst 33258 staining assay. The results indicated that KD showed antiangiogenic activity via inhibitory effects on cell proliferation, cell migration, cell adhesion, and tube formation. ROS levels were down-regulated and catalase activity was up-regulated after treatment with KD. The cell cycle was arrested at the G2/M phase, while no apoptosis was observed using the Hoechst 33258 staining assay or following the flow cytometric analysis of the sub-G1 proportion. The antiangiogenic properties of KD, in combination with its anti-proliferative effect and ability to induce cell cycle arrest without inducing apoptosis, make it a good candidate for development as antitumor agent. However, further studies are essential to elucidate its mechanism of action.