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目的:研究冬凌草甲素抑制血管生成的活性及其作用机制。方法:采用体外和体内两种模式,研究冬凌草甲素在血管生成方面的抑制效应。通过体外培养心脏微血管内皮细胞,噻唑蓝(MTT)比色法检测细胞的活力,酶联免疫吸附(ELISA)法测定血管内皮生长因子(VEGF)的含量;体内选择斑马鱼(Fli1-GFP)生物模式,观察冬凌草甲素对胚胎期血管生成和成鱼期血管损伤后再生的影响,并采用相对荧光定量PCR法检测VEGF通路上主要相关基因VEGFA,血管内皮细胞生长因子受体(VEGFR)2,VEGFR3的表达量。结果:冬凌草甲素具有抑制血管生成活性的作用,可抑制体外内皮细胞活力,其半抑制浓度(IC50)8.04 mg·L~(-1),可使细胞血清中VEGF的表达量明显下降;可抑制斑马鱼胚胎期体节间血管生成,且抑制成鱼期血管损伤后再生,可能通过降低体内VEGFA,VEGFR2,VEGFR3基因的表达量从而抑制血管生成。结论:冬凌草甲素可有效抑制血管生成,为其抗肿瘤血管生成治疗提供了重要的科学依据。
Objective: To investigate the inhibitory effect of oridonin on angiogenesis and its mechanism. Methods: The inhibitory effects of oridonin on angiogenesis were studied in vitro and in vivo. Cardiac microvascular endothelial cells were cultured in vitro, the viability of cells was detected by MTT colorimetric assay, and the content of vascular endothelial growth factor (VEGF) was determined by enzyme-linked immunosorbent assay (ELISA). In vivo, zebrafish (Fli1-GFP) The effects of oridonin on the angiogenesis in embryonic stage and the regeneration after vascular injury in adult fish were observed. Relative expression of VEGFA, vascular endothelial growth factor receptor (VEGFR) 2, the expression of VEGFR3. Results: Oridonin could inhibit the activity of angiogenesis and inhibit the activity of endothelial cells in vitro. The IC50 of 8.04 mg · L -1 could inhibit the expression of VEGF in serum significantly ; It can inhibit the internode angiogenesis in the zebrafish embryos and inhibit the regeneration after vascular injury in adult fish. It may inhibit the angiogenesis by reducing the expression of VEGFA, VEGFR2 and VEGFR3 in vivo. Conclusion: Oridonin can effectively inhibit angiogenesis and provide an important scientific basis for its antiangiogenic therapy.