红花全提取物在体外人微血管内皮细胞和体内斑马鱼中的促血管生成作用。

Pro-angiogenic effects of Carthami Flos whole extract in human microvascular endothelial cells in vitro and in zebrafish in vivo.

作者信息

Zhou Xuelin, Siu Wing-Sum, Fung Chak-Hei, Cheng Ling, Wong Chun-Wai, Zhang Cheng, Liu Cheuk-Lun, Kwok Hin-Fai, Lau Ching-Po, Wat Elaine, Lau Clara Bik-San, Leung Ping-Chung, Ko Chun-Hay, Hung Leung-Kim

机构信息

Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong Special Administrative Region; State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong Special Administrative Region; Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, Guangdong Province, China.

出版信息

Phytomedicine. 2014 Sep 25;21(11):1256-63. doi: 10.1016/j.phymed.2014.06.010. Epub 2014 Jul 25.

DOI:10.1016/j.phymed.2014.06.010

PMID:25172787

Abstract

AIM

Carthami Flos (CF) is a Chinese herb traditionally used for cardiovascular disease and bone injury in China with pharmacological effects on improving blood circulation. The aim of this study was to investigate the angiogenic potential of CF whole extract (extracted by boiling with water, followed by ethanol) and the underlying mechanisms in human microvascular endothelial cells (HMEC-1) in vitro and in transgenic TG(fli1:EGFP)(y1)/+(AB) zebrafish with transgenic endothelial cells expressing EGFP (Enhanced Green Fluorescent Protein) in vivo.

METHODS

Effects of CF whole extract on cell proliferation, migration and tube formation in HMEC-1 cells in vitro were detected by MTT assay, wound healing assay and tube formation assay. Its angiogenic effect in zebrafish was investigated by monitoring the sprout number in the sub-intestinal vessel (SIV), and the underlying mechanisms were tested by quantitative real-time PCR.

RESULTS

CF whole extract increased cell proliferation, migration and tube formation in vitro in HMEC-1 cells. Its angiogenic effect was also confirmed in vivo in zebrafish by increasing the sprout number in the SIV. As determined by quantitative real-time PCR, CF whole extract up-regulated the expression of angiogenesis-related genes in zebrafish, including angiogenic and its associated growth factors and receptors (e.g. IGF1, CTGF, NRP2, and VEGFR3), transcription factor (e.g. HIF1A), matrix degradation and endothelial cell migration-related factors (e.g. MMP2, MMP9, TIMP2, PLG and PLAU), cell adhesion molecules (e.g. ITGAV, ITGB3, beta-catenin and PECAM1), tubule formation factors (e.g. ANGPT1, TIE-2, PDGFR-B, CDH5, S1PR1, FGF2, Shh, and TGFRB1), and blood vessel maturation/formation factor (e.g. Ephrin B2).

CONCLUSIONS

CF whole extract increased angiogenesis in HMEC-1 cells in vitro and in zebrafish in vivo with multiple mechanisms.

摘要

目的

红花是一种传统上用于治疗中国心血管疾病和骨损伤的中药，具有改善血液循环的药理作用。本研究旨在探讨红花全提取物（经水煮后用乙醇提取）在体外人微血管内皮细胞（HMEC-1）和体内转基因TG(fli1:EGFP)(y1)/+(AB)斑马鱼（其转基因内皮细胞表达增强型绿色荧光蛋白）中的血管生成潜力及潜在机制。

方法

采用MTT法、伤口愈合试验和管腔形成试验检测红花全提取物对体外HMEC-1细胞增殖、迁移和管腔形成的影响。通过监测肠下血管（SIV）中的芽生数量来研究其在斑马鱼中的血管生成作用，并通过定量实时PCR检测潜在机制。

结果

红花全提取物在体外可促进HMEC-1细胞的增殖、迁移和管腔形成。在体内斑马鱼中，通过增加SIV中的芽生数量也证实了其血管生成作用。定量实时PCR检测结果显示，红花全提取物上调了斑马鱼中血管生成相关基因的表达，包括血管生成及其相关生长因子和受体（如IGF1、CTGF、NRP2和VEGFR3）、转录因子（如HIF1A）、基质降解和内皮细胞迁移相关因子（如MMP2、MMP9、TIMP2、PLG和PLAU）、细胞黏附分子（如ITGAV、ITGB3、β-连环蛋白和PECAM1）、小管形成因子（如ANGPT1、TIE-2、PDGFR-B、CDH5、S1PR1、FGF2、Shh和TGFRB1）以及血管成熟/形成因子（如Ephrin B2）。