叶酸挽救乙醇致斑马鱼胚胎心脏和流出道发育异常  

The folic acid rescue of ethanol-induced heart and outflow tract malformations in zebrafish

作　　者：孙淑娜[1,2] 姚泽忠 蒋璆[3] 桂永浩[1] 

机构地区：[1]复旦大学附属儿科医院心内科,上海201102 [2]上海市闵行区中心医院儿科,上海201199 [3]上海市心血管病研究所,上海200032

出　　处：《复旦学报（医学版）》2016年第2期135-141,共7页Fudan University Journal of Medical Sciences

基　　金：上海市卫生局青年科研基金(20124y086);上海市出生缺陷防治重点实验室开放课题~~

摘　　要：目的观察乙醇对斑马鱼胚胎心脏和流出道发育的干扰作用,并初步探讨叶酸挽救乙醇所致心脏和流出道发育异常的最佳时段和可能机制。方法在斑马鱼胚胎发育不同时段予以乙醇处理,观察胚胎的发育异常情况。将受精后7~12 h被予以乙醇组定义为乙醇处理组。在乙醇处理组胚胎的发育不同时段给予外源性叶酸进行叶酸挽救实验,观察各叶酸干预组胚胎的发育状况,探寻叶酸挽救的最佳时段。将受精后7~12 h给予叶酸组定义为叶酸挽救组。观察并统计乙醇处理组以及叶酸挽救组的胚胎发育异常百分比、存活百分比、心脏和流出道形态及其异常百分比、心率、心室收缩指数等指标来评估胚胎的发育状况。利用荧光显微造影的方法探查心脏流出道形态,采用胚胎整体原位杂交和Real-time PCR方法检测胚胎bmp2b、tbx1的表达情况。结果乙醇可干扰胚胎发育,乙醇处理组胚胎存在明显心脏和流出道形态异常以及心功能损害,在胚胎发育早期乙醇的干扰作用最明显。给予外源性叶酸能挽救乙醇导致的胚胎发育异常,在胚胎受精后7~12 h叶酸挽救作用最显著。乙醇处理组胚胎心脏和流出道的bmp2b、tbx1表达下降;予以叶酸干预后,乙醇处理组胚胎bmp2b和tbx1表达水平上调。结论叶酸能有效挽救乙醇导致的心脏和流出道发育异常,其机制可能是上调乙醇处理组胚胎bmp2b、tbx1的表达水平。Objective To observe ethanol-induced cardiac phenotypes and identify ethanol-sensitive stages during embryonic development,and to explore the optimal period during which the administration of folic acid can effectively rescue the ethanol-induced heart and outflow tract（OFT）defects and its underlying mechanisms. Methods Ethanol was administrated to zebrafish embryos in a series of developmental stages（7-12 h）and the abnormal heart and OFT phenotypes were observed after ethanol exposure.The embryos which were given ethanol at 7-12 hpf was defined as ethanol treated group.Administrating folic acid to ethanol-treated zebrafish embryos in different stages（7-12h）was conducted to rescue the abnormal cardiac defects.The ethanol-treated embryos which were given folic acid at 7-12 hpf was defined as folic acid rescue group.The survival percentage,the malformation percentage,the heart rate and the ventricular shortening fraction（VSF）were recorded.The OFT phenotypes were evaluated using fluorescein micro-angiography.Whole-mount in situ hybridization and Real-time PCR were conducted to analyze the expression of bmp2 b and tbx1.Results Ethanol exposure produced heart and OFT defects,including the abnormal cardiac shapes,the hypogenesis of OFT and the reduced heart rate and VSF.The teratogenic effect of ethanol was observed most obviously at the early embryonic development.Supplementation of folic acid at 7-12 hours post-fertilization partially rescued the developmental defects in ethanol-induced zebrafish embryos.The expressions of bmp2 b and tbx1 were reduced in ethanol-treated group,and they were partially recovered after administration of folic acid. Conclusions Folic acid supplementation can effectively rescue ethanol-induced heart OFT defects,possibly by enhancing the expressions of bmp2 b and tbx1.

关 键 词：叶酸 乙醇 心脏 流出道 斑马鱼 bmp2b tbx1 

分 类 号：R725.4[医药卫生—儿科] R394.1[医药卫生—临床医学]