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藏茵陈水提物对斑马鱼新生血管的抑制效果及调控机制
刘源,张莉敏,熊飞,由凤鸣,蔡懿,陈婧,冯阳,万雪梅
0
(成都中医药大学临床医学院·附属医院;成都中医药大学肿瘤研究所)
摘要:
【摘要】 目的 探讨藏茵陈水提物对斑马鱼新生血管的抑制效果及调控机制。 方法 选取TG 品系的 fli1aEGFP 转基因斑马鱼为实验动物,收集90个22hpf发育正常的斑马鱼胚胎,分为空白对照组、阳性对照组、藏茵陈水提物组,每组各30例。空白对照组予以0.1%二甲基亚砜(DMSO)、阳性对照组予以瓦他拉尼(工作浓度为 5μg/ml)、藏茵陈水提物组予以藏茵陈水提取物(工作浓度为 200μg/ml)。在体式荧光显微镜下观察藏茵陈提取物处理的鱼胚血管表型并拍照,分析各样品对斑马鱼体节间血管(ISVs)生成的影响。利用实时荧光定量 PCR 技术检测处理后鱼胚的ptprb、nr2f1a、VEcadherin、S1P1、CD146、pik3r2、netrin1a、fgfr1a、fgfr2、fgfr3、fgfr4基因的表达情况。 结果 与空白对照组及阳性对照组相比较,藏茵陈水提取物组鱼胚ISVs变薄,背主动脉出芽减少,显示出抑制斑马鱼血管生成的作用(P<0.05)。藏茵陈水提物组的nr2f1a基因出现下调,但与空白对照组及阳性对照组相比较差异无统计学意义(P>005),ptprb基因出现下调,差异有统计学意义(P<0.0001),VEcadherin、S1P1、CD146、pik3r2、netrin1a、fgfr1a、fgfr2、fgfr3、fgfr4基因出现上调,但与空白对照组及阳性对照组比较差异无统计学意义(P>0.05)。 结论 藏茵陈水提取物有抑制斑马鱼新生血管的作用,其机制与下调ptprb的基因表达有关。
关键词:  斑马鱼  新生血管  藏茵陈  ptp rb基因  抑制  调控
DOI:
基金项目:四川省科技支撑计划项目
Inhibitory effect of aqueous extract of Capillary wormwood herb on neovascularization of zebrafish and its regulatory mechanism
LIU Yuan,ZHANG Limin,XIONG Fei,YOU Fengming,CAI Yi,CHEN Jing,FENG Yang,WAN Xuemei
(Clinical Medical School/Teaching Hospital, Chengdu University of Traditional Chinese Medicine;Institution of Cancer research, Chengdu University of Traditional Chinese Medicine)
Abstract:
【Abstract】 Objective To explore the inhibitory effect and regulatory mechanism of water extract of Capillary wormwood herb on zebrafish neovascularization. Methods 90 embryos of 22 HPF normal development zebrafish were collected from fli1a EGFP transgenic zebrafish of TG strain. They were divided into blank control group, positive control group and Capillary wormwood herb water extract group. The blank control group was given 0.1% DMSO, the positive control group was given 5 μg/ml vatarani, the water extract group was given 200 μg/ml. The vascular phenotypes of zebrafish embryos treated with the extract of Artemisia Capillaria were observed and photographed under the body fluorescence microscope, and the effects of the samples on the formation of the internode vessels (ISVs) of zebrafish were analyzed. The expression of PTP Rb, nr2f1a, VE cadherin, S1P1, CD146, pik3r2, netrin1a, fgfr1a, FGFR2, FGFR3, FGFR4 genes in the treated fish embryos was detected by realtime fluorescence quantitative PCR. Results Compared with the blank control group and the positive control group, the ISVs of the fish embryo in the water extract group of Artemisia Capillaria became thinner and the sprouting of the dorsal aorta decreased, showing the inhibition effect on the angiogenesis of zebrafish (P<005). The nr2f1a gene of the water extract group was down regulated, but there was no significant difference compared with the blank control group and the positive control group (P>005), PTP Rb gene was down regulated, the difference was statistically significant (P<00001). The expression of VEcadherin, S1P1, CD146, pik3r2, netrin1a, fgfr1a, fgfr2, fgfr3 and fr4 increased, But there was no significant difference between the two groups (P>005). Conclusion The water extract of Capillary wormwood herb can inhibit the neovascularization of zebrafish, and its mechanism is related to the downregulation of PTP Rb gene expression.
Key words:  Zebrafish  Neovascularization  Capillary wormwood herb  ptp rb gene  Restrain  Regulation

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