天然产物抗血管生成的体内筛选及脱氧苏木酮B 7,4'-二甲醚的抗血管生成活性机制

In vivo Screening of Natural Products Against Angiogenesis and Mechanisms of Anti-Angiogenic Activity of Deoxysappanone B 7,4'-Dimethyl Ether.

作者信息

Chen Kan, Fan Yuqi, Gu Jun, Han Zhihua, Zeng Huasu, Mao Chengyu, Wang Changqian

机构信息

Department of Cardiology, Shanghai Ninth People's Hospital Affiliated Shanghai Jiaotong University School of Medicine, Shanghai 200011, People's Republic of China.

出版信息

Drug Des Devel Ther. 2020 Jul 30;14:3069-3078. doi: 10.2147/DDDT.S252681. eCollection 2020.

INTRODUCTION

The aim of this study was to screen the leading compounds of natural origin with anti-angiogenic potential and to investigate their anti-angiogenic mechanism preliminarily.

MATERIALS AND METHODS

An initial screening of 240 compounds from the Natural Products Collection of MicroSource was performed using the transgenic zebrafish strain . The zebrafish embryos at 24 h post-fertilization were exposed to the natural compounds for an additional 24 h; then, morphological changes in the intersegmental vessels (ISVs) were observed and quantified under a fluorescence microscope. The expression profiles of angiogenesis-related genes in the zebrafish embryos were detected using quantitative real-time PCR.

RESULTS

Five compounds were identified with potential anti-angiogenic activity on the zebrafish embryogenesis. Among them, deoxysappanone B 7.4'-dimethyl ether (Deox B 7,4) showed anti-angiogenic activity on the formation of ISVs in a dose-dependent manner. The inhibition of ISV formation reached up to 99.64% at 5 μM Deox B 7,4. The expression of delta-like ligand 4 (), hes-related family basic helix-loop-helix transcription factor with YRPW motif 2, ephrin B2, fibroblast growth factor receptor () 3, cyclooxygenase-2, protein tyrosine phosphatase, receptor type B (), phosphoinositide-3-kinase regulatory subunit 2, slit guidance ligand () 2, , roundabout guidance receptor () 1, , and were down-regulated, while vascular endothelial growth factor receptor-2, fgfr 1, and matrix metallopeptidase 9 were up-regulated in the zebrafish embryos treated with Deox B 7,4.

CONCLUSION

Deox B 7,4 has a therapeutic potential for the treatment of angiogenesis-dependent diseases and may exert anti-angiogenic activities by suppressing the slit2/robo1/2, slit3/robo4, cox2/ptp-rb/pik3r2, and dll4/hey2/efnb2a signaling pathways as well as activation of vegfr-2/fgfr1/mmp9.

引言

本研究旨在筛选具有抗血管生成潜力的天然来源先导化合物，并初步探究其抗血管生成机制。

材料与方法

使用转基因斑马鱼品系对MicroSource天然产物库中的240种化合物进行初步筛选。将受精后24小时的斑马鱼胚胎再暴露于天然化合物中24小时；然后，在荧光显微镜下观察并量化节间血管（ISV）的形态变化。使用定量实时PCR检测斑马鱼胚胎中血管生成相关基因的表达谱。

结果

鉴定出5种对斑马鱼胚胎发育具有潜在抗血管生成活性的化合物。其中，脱氧苏木酮B 7,4'-二甲醚（Deox B 7,4）对ISV的形成具有剂量依赖性的抗血管生成活性。在5 μM Deox B 7,4时，ISV形成的抑制率高达99.64%。在经Deox B 7,4处理的斑马鱼胚胎中，类Notch配体4（Delta-like ligand 4，Dll4）、含YRPW基序的hes相关家族碱性螺旋-环-螺旋转录因子2（hes-related family basic helix-loop-helix transcription factor with YRPW motif 2，Hey2）、ephrin B2、成纤维细胞生长因子受体（fibroblast growth factor receptor，Fgfr）3、环氧化酶-2（cyclooxygenase-2，Cox2）、蛋白酪氨酸磷酸酶受体B型（protein tyrosine phosphatase, receptor type B，Ptp-Rb）、磷脂酰肌醇-3-激酶调节亚基2（phosphoinositide-3-kinase regulatory subunit 2，Pik3r2）、缝隙引导配体（slit guidance ligand，Slit）2、Roundabout引导受体（roundabout guidance receptor，Robo）1、Robo4以及Hey1的表达下调，而血管内皮生长因子受体-2（vascular endothelial growth factor receptor-2，Vegfr-2）、Fgfr 1和基质金属肽酶9（matrix metallopeptidase 9，Mmp9）的表达上调。