地塞米松诱导新西兰兔先天性腭裂的非靶向代谢组学研究

Untargeted metabolomics study of dexamethasone-induced congenital cleft palate in New Zealand rabbits

目的应用非靶向代谢组学技术比较地塞米松干预后的腭裂胚胎、非腭裂胚胎与正常胚胎在腭突发育期间胎盘组织的差异。方法将12只孕兔(40周龄,体质量4.5~5.0 kg的雌性新西兰兔)按照随机数表法分为地塞米松组(8只)和对照组(4只),在妊娠第13到16天每日分别给两组孕兔后腿四头肌肌肉注射1次1.0 mg地塞米松和1 ml生理盐水,并于妊娠第21天处死孕兔,剖腹取其子代,按照子代腭部表型分为3组,分别为地塞米松腭裂组、地塞米松非腭裂组(经地塞米松未诱导腭裂)和正常对照组,再收集胎盘组织样本。应用液相色谱—三重串联四级杆液质联用仪获取胚胎胎盘组织样本的相应图谱,主成分分析法描绘3组胚胎胎盘组织代谢产物在胚胎腭突发育阶段的代谢差异。结果地塞米松腭裂组、地塞米松非腭裂组和正常对照组之间存在明显的代谢差异,差异代谢物共133种(VIP>1,P<0.05),涉及的重要代谢通路包括维生素B6代谢、赖氨酸代谢、精氨酸合成代谢、半乳糖代谢。正常对照组、地塞米松非腭裂组和地塞米松腭裂组中的维生素B6(分别为0.960±0.249、0.856±0.368、1.319±0.322)、半乳糖(分别为0.888±0.171、1.033±0.182、1.127±0.127)、赖氨酸(分别为1.551±0.924、1.789±1.435、0.541±0.424)和尿素(分别为0.743±0.142、1.137±0.301、1.171±0.457)差异均有统计学意义(F=5.90,P=0.008;F=5.59,P=0.009;F=4.26,P=0.025;F=5.29,P=0.012)。结论基于胎盘组织代谢物建立的统计模型表明,3组胚胎胎盘组织的代谢物表达具有差异;地塞米松诱导腭裂形成可能与维生素B6代谢、赖氨酸代谢、精氨酸合成代谢、半乳糖代谢高度相关。

Objective To investigate the metabolic disorders in placental tissues of dexamethasone induced cleft palate mode.Methods Twelve pregnant rabbits were randomly divided into dexamethasone group(experimental group,8)and saline control group(4),and a certain amount of dexamethasone and saline were administered intramuscularly to the experimental and control groups respectively from embryonic days(ED)13 to 16,and placental tissue samples were collected on day 21 of gestation.The corresponding profiles of the embryonic placental tissue samples were obtained by liquid chromatography-triple tandem quadrupole(LC-MS),and the metabolites of the embryonic placental tissues were characterized by principal component analysis among the dexamethasone-treated group with cleft palate(D-CP group),the dexamethasone-treated group without cleft palate(D-NCP group)and the control group.Results There were significant metabolic differences among the D-CP group,D-NCP group and control group,with a total of 133 differential metabolites(VIP>1,P<0.05)involving in important metabolic pathways including vitamin B6 metabolism,lysine metabolism,arginine anabolic metabolism,and galactose metabolism.The four metabolites,vitamin B6,galactose,lysine and urea,differed among the three groups(P<0.05).There were significant differences in vitamin B6(0.960±0.249,0.856±0.368,1.319±0.322),galactose(0.888±0.171,1.033±0.182,1.127±0.127),lysine(1.551±0.924,1.789±1.435,0.541±0.424)and urea(0.743±0.142,1.137±0.301,1.171±0.457,respectively)levels among control group,D-NCP group and D-CP group(F=5.90,P=0.008;F=5.59,P=0.009;F=4.26,P=0.025;F=5.29,P=0.012).Conclusions The results indicated that dexamethasone induced cleft palate may be highly correlated with metabolic disorders including vitamin B6 metabolism,lysine metabolism,arginine anabolic metabolism and galactose metabolism.