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Angiogenesis (angiogenesis) is the existing blood vessels (such as capillaries and venules) produced by budding or fission process of new blood vessels. Embryogenesis, the menstrual cycle, wound healing and physiological angiogenesis; diabetes, proliferative retinopathy, atherosclerosis, psoriasis, rheumatoid arthritis, cancer and other diseases are associated with pathological angiogenesis. Increase in tumor volume and tumor cell metastasis depend on angiogenesis, thus inhibiting tumor angiogenesis has become the hot research and development of anticancer drugs, and anti-angiogenic therapy has become a new strategy for cancer therapy. Numerous studies show that flavonoids affect the activity of various enzymes. In addition to having antibacterial, anti-inflammatory, anti-mutagenic, blood pressure, detoxification, sedation and other effects, the anti-oxidation, anti-cancer, anti-cancer, inhibit angiogenesis also has a significant effect. Kushecarpin D (KD) is extracted from Sophora novel flavonoids. Its efficacy and pharmacological been reported. In this experiment, cultured human umbilical vein endothelial cells (ECV304) as a model to study the flavescens flavonoids KD antiangiogenic effects and related mechanisms, in order to provide for the further development and utilization of experimental evidence. The experiment was divided into in vitro anti-angiogenic effect and mechanism of two related parts. The first part was evaluated by SRB assay KD effects on cell proliferation, ranked by trypan blue staining of the KD cytotoxicity through cell adhesion, cell migration, tube-like structure formation angiogenesis benchmark the KD antiangiogenic generation effect. The second part of cell cycle by flow cytometry using DCFH-DA fluorescence assay ROS generation, using ammonium molybdate colorimetric determination of catalase activity, as well as Hoechst 33258 staining apoptosis. The results showed that: 1, KD inhibit angiogenesis ECV304 by 10,20,30,40,50 μg / ml KD for 12 h, cell proliferation was detected by SRB assay was found to be significantly inhibited cell growth, but 30,40,50 μg / ml The inhibition rate was essentially flat. Trypan blue staining row KD results show inhibition of cell proliferation is not achieved by killing cells. KD weak effect on cell migration, drug concentration of 40μg/ml, the cell migration was inhibited significantly (p lt; 0.05). Since the beginning of drug concentration 20μg/ml, ECV304 cell adhesion that was inhibited in a dose-dependent. The results showed that the formation of tube-like structure, 10μg/ml KD tube-like structure that is formed showing a very significant inhibition, the inhibition was dose-dependent. This indicates KD on ECV304 cell proliferation, migration, adhesion, tube-like structure formation angiogenesis indicators have shown inhibition. 2, KD antiangiogenic effect may be related to cell cycle arrest at the G2 / M phase and suppress the generation of intracellular H2O2. 40μg/ml, 50μg/ml KD treated cells, the cell cycle was arrested at G2 / M phase, but did not induce apoptosis. Add KD treatment, the cells were significantly lower H2O2 content, which KD consistent with inhibition of angiogenesis. Show KD antiangiogenic effect may be related cell cycle arrest at G2 / M phase, inhibit the generation of intracellular H2O2.
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