代谢组学评估胚胎发育潜能的研究进展

体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET,简称IVF)技术已成为治疗不孕症的重要手段,在不降低临床妊娠率的同时减少多胎妊娠率是亟待解决的重要问题.常规形态学评分筛选胚胎的方法往往主观性太强,而胚胎植入前遗传学筛查技术又具有侵入性.因此,寻找一种非侵入...     

基于液相质谱技术探讨阿霉素诱导扩张型心肌病潜在血浆标志物

目的:扩张性心肌病是应用阿霉素治疗肿瘤的严重副作用之一,致死率可达50％.通过建立阿霉素扩张型心肌病动物模型对比健康动物,寻找该疾病的生物标志物.方法:采用阿霉素注射法制备扩张型心肌病模型,造模后收集模型组与对照组大鼠的血浆与心肌组织,应用液相质谱技术检测血浆及心肌代谢物,选择共同存在于血浆及心肌组织中的代谢物,并应用...     

Metabolomic method to detect a metabolite corona on amino-functionalized polystyrene nanoparticles

Abstract

Protein coronas on nanoparticles (NPs) affect their physicochemical properties, cellular uptake, and toxicity, and have been described extensively. To date, studies of the occurrence of small molecule (metabolite) coronas are limited. We sought to determine whether a metabolite corona forms on NPs, using high-sensitivity metabolomics combined with a model system for freshwater ecotoxicology (Daphnia magna feeding on Chlorella vulgaris). Using amino-functionalized polystyrene NPs (NH₂-pNPs), we showed the impact of this material on Daphnia feeding to provide a rationale for the detailed molecular investigations. We then employed a targeted LC-MS/MS approach for sodium dodecyl sulfate (SDS) as an analog to signaling molecules known to occur in our freshwater model system and optimized a corona extraction method for this representative metabolite. Next, we performed an untargeted discovery-based metabolomics study – using high-sensitivity nanoelectrospray direct infusion mass spectrometry (DIMS) – to enable an unbiased assessment of the metabolite corona of NH₂-pNPs in the freshwater model system. Our results demonstrate that SDS was successfully recovered from NH₂-pNPs, confirming that the extraction protocol was fit-for-purpose. Untargeted DIMS metabolomics reproducibly detected 100?s of small molecule peaks extracted from NH₂-pNPs exposed to conditioned media from the D. magna–C. vulgaris model system. Attempts to annotate these extracted metabolites, including by using van Krevelen and Kendrick Mass Defect plots, indicate a diverse range of metabolites that were not clustered into any particular class. Overall we demonstrate the existence of an ecologically relevant metabolite corona on the surface of NPs through application of a high-sensitivity, untargeted mass spectrometry metabolomics workflow.     

Evolution of the Ergot Alkaloid Biosynthetic Gene Cluster Results in Divergent Mycotoxin Profiles in Claviceps purpurea Sclerotia

Research into ergot alkaloid production in major cereal cash crops is crucial for furthering our understanding of the potential toxicological impacts of Claviceps purpurea upon Canadian agriculture and to ensure consumer safety. An untargeted metabolomics approach profiling extracts of C. purpurea sclerotia from four different grain crops separated the C. purpurea strains into two distinct metabolomic classes based on ergot alkaloid content. Variances in C. purpurea alkaloid profiles were correlated to genetic differences within the lpsA gene of the ergot alkaloid biosynthetic gene cluster from previously published genomes and from newly sequenced, long-read genome assemblies of Canadian strains. Based on gene cluster composition and unique polymorphisms, we hypothesize that the alkaloid content of C. purpurea sclerotia is currently undergoing adaptation. The patterns of lpsA gene diversity described in this small subset of Canadian strains provides a remarkable framework for understanding accelerated evolution of ergot alkaloid production in Claviceps purpurea.     

基于代谢组学的芩百清肺浓缩丸治疗支原体肺炎小鼠的作用机制研究

目的研究芩百清肺浓缩丸治疗支原体肺炎的作用机制。方法采用超高效液相色谱与串联四级杆飞行时间质谱(UPLC-Q-TOF-MS)联用技术的代谢组学方法,分析肺炎支原体感染小鼠肺组织中内源性化合物的变化。采用Progenisis QI软件进行色谱峰识别及匹配,并采用主成分分析(PCA)和偏最小二乘-判别分析(PLS-DA)对获得数据进行降维,通过分析对不同组间分离贡献度较大(VIP1,P0.05)化合物的串联质谱数据,经HMDB等数据库检索,确定潜在生物标志物。结果从肺组织中共鉴定出视黄醛、全反式维甲酸、焦谷氨酸、白三烯C4、维生素A、花生四烯酸、前列腺素I2等20个潜在生物标志物。与模型组比较,芩百清肺浓缩丸对20个潜在生物标志物均具有回调作用。结论芩百清肺浓缩丸通过影响视黄醇代谢、亚油酸代谢、花生四烯酸代谢等通路发挥治疗支原体肺炎的作用。     

左卡尼汀缓解顺铂造成急性肾损伤的血清代谢组学研究

目的 利用血清代谢组学探究顺铂诱导小鼠急性肾损伤的特异性变量,同时评价左卡尼汀的干预作用。 方法 将19只小鼠分为正常对照组、模型组和干预组,适应3 d后,对干预组给予左卡尼汀(400 mg/kg,ip)干预,2 d后给予模型组和干预组顺铂(20 mg/kg,ip)造模,每天称量各组小鼠的体质量,2 d后取小鼠血清进行LC-MS分析,结合模式识别分析各组间代谢组差异,并评价左卡尼汀的干预作用。 结果 代谢组学分析共鉴别28个差异代谢物,顺铂诱导的急性肾损伤主要涉及磷脂类、氨基酸类和脂肪酸类代谢途径的改变,而左卡尼汀有改善作用。 结论 左卡尼汀可改善顺铂诱导的急性肾损伤,其机制可能是通过调控色氨酸代谢、谷氨酸代谢和能量代谢,从而减缓急性肾损伤的疾病进程。     

Effects of Low Doses of Bisphenol A on the Metabolome of Perinatally Exposed CD-1 Mice

Background: Bisphenol A (BPA) is a well-known endocrine disruptor used to manufacture polycarbonate plastics and epoxy resins. Exposure of pregnant rodents to low doses of BPA results in pleiotropic effects in their offspring.Objective: We used metabolomics—a method for determining metabolic changes in response to nutritional, pharmacological, or toxic stimuli—to examine metabolic shifts induced in vivo by perinatal exposure to low doses of BPA in CD-1 mice.Methods: Male offspring born to pregnant CD-1 mice that were exposed to vehicle or to 0.025, 0.25, or 25 µg BPA/kg body weight/day, from gestation day 8 through day 16 of lactation, were examined on postnatal day (PND) 2 or PND21. Aqueous extracts of newborns (PND2, whole animal) and of livers, brains, and serum samples from PND21 pups were submitted to ¹H nuclear magnetic resonance spectroscopy. Data were analyzed using partial least squares discriminant analysis.Results: Examination of endogenous metabolic fingerprints revealed remarkable discrimination in whole extracts of the four PND2 newborn treatment groups, strongly suggesting changes in the global metabolism. Furthermore, statistical analyses of liver, serum, and brain samples collected on PND21 successfully discriminated among treatment groups. Variations in glucose, pyruvate, some amino acids, and neurotransmitters (γ-aminobutyric acid and glutamate) were identified.Conclusions: Low doses of BPA disrupt global metabolism, including energy metabolism and brain function, in perinatally exposed CD-1 mouse pups. Metabolomics can be used to highlight the effects of low doses of endocrine disruptors by linking perinatal exposure to changes in global metabolism.     

Metabolomics as a powerful tool for molecular quality assessment of the fish Sparus aurata

The molecular profiles of perchloric acid solutions extracted from the flesh of Sparus aurata fish specimens, produced according to different aquaculture systems, have been investigated. The (1)H-NMR spectra of aqueous extracts are indicative of differences in the metabolite content of fish reared under different conditions that are already distinguishable at their capture, and substantially maintain the same differences in their molecular profiles after sixteen days of storage under ice. The fish metabolic profiles are studied by top-down chemometric analysis. The results of this exploratory investigation show that the fish metabolome accurately reflects the rearing conditions. The level of many metabolites co-vary with the rearing conditions and a few metabolites are quantified including glycogen (stress indicator), histidine, alanine and glycine which all display significant changes dependent on the aquaculture system and on the storage times.     

From a non-targeted metabolomics approach to a targeted biomarkers strategy to highlight testosterone abuse in equine. Illustration of a methodological transfer between platforms and laboratories

In order to overcome the challenge associated with the screening of Anabolic-Androgenic Steroids abuses in animal competitions, a non-targeted liquid chromatography coupled to high resolution mass spectrometry based metabolomics approach was implemented on equine urine samples to highlight potential biomarkers associated with the administration of such compounds, using testosterone esters as model steroids. A statistical model relying on four potential biomarkers intensity could be defined to predict the status of the samples. With a routine application perspective, the monitoring of the highlighted potential biomarkers was first transferred into high-throughput liquid chromatography-selected reaction monitoring (LC-SRM). The model's performances and robustness of the approach were preserved and providing a first demonstration of metabolomics-based biomarkers integration within a targeted workflow using common benchtop MS instrumentation. In addition, with a view to the widespread implementation of such biomarker-based tools, we have transferred the method to a second laboratory with similar instrumentation. This proof of concept allows the development and application of biomarker-based strategies to meet current doping control needs.     

基于非靶向代谢组学的促乳腺癌曲妥珠单抗耐药 代谢产物分析

摘 要 目的：探究人类表皮生长因子受体2（HER2）阳性乳腺癌曲妥珠单抗（trastuzumab）耐药细胞（HCC1954）和敏感细胞（SK-BR-3）之间的代谢物差异。方法：利用超高效液相色谱-串联质谱（UHPLC-MS/MS）非靶向代谢组学方法进行代谢物的鉴定与定量,结合多元统计方法主成分分析（PCA）和正交偏最小二乘-判别分析（OPLS-DA）以筛选差异代谢物,并对差异代谢物进行KEGG通路富集分析。结果：在正负离子模式下鉴定筛选出15种显著性变化差异代谢物,在曲妥珠单抗耐药细胞中丙酰肉碱和谷胱甘肽等7种代谢物上调,肉豆蔻酸和D-葡萄糖酸等8种代谢物下调。KEGG通路富集分析发现,9条代谢通路具有统计学意义（P<0.05）,其中氮代谢和谷氨酰胺与谷氨酸代谢最为显著,P值为0.001 3,还发现谷氨酸和谷氨酰胺均在这两条通路中发挥作用。结论：本研究初步揭示了HER2阳性乳腺癌曲妥珠单抗耐药和敏感细胞之间的代谢物差异,为HER2阳性乳腺癌细胞曲妥珠单抗诊断标志物进一步研究提供了代谢组学的数据支持。