<Superscript>1</Superscript>H-NMR metabonomics analysis of sera differentiates between mammary tumor-bearing mice and healthy controls

Global analysis of ¹H-NMR spectra of serum is an appealing approach for the rapid detection of cancer. To evaluate the usefulness of this method in distinguishing between mammary tumor-bearing mice and healthy controls, we conducted ¹H-NMR metabonomic analyses on serum samples obtained from the following: 10 mice inoculated with a highly-metastatic mammary carcinoma cell line, 10 mice inoculated with a “normally” metastatic mammary carcinoma cell line, and 10 healthy controls. Following standard spectral processing and subsequent data reduction, we applied unsupervised Principal Component Analysis (PCA) to determine if unique metabolic fingerprints for different categories of metastatic breast cancer in serum exist. The PCA method correctly separated sera of tumor-bearing mice from that of normal healthy controls, as shown using the scores plot which indicated that sera classes from tumor-bearing mice did not share multivariate space with that from healthy controls. In addition, this technique was capable of distinguishing between classes of varying metastatic ability in this system. Metabolites apparently responsible for separation between diseased and healthy mice include lactate, taurine, choline, and sugar moieties. Results of this study suggest that ¹H-NMR spectra of mouse serum analyzed using PCA statistical methods indicate separation of tumor-bearing mice from healthy normal controls, justifying further study of the use of ¹H-NMR metabonomics for cancer detection using serum.     

汉黄芩苷通过调节抗氧化基因表达和机体代谢延长果蝇寿命

为探究汉黄芩苷是否具有抗衰老作用,本研究以果蝇为模型,考察汉黄芩苷对果蝇自然寿命的影响。采用RT-PCR和UPLC-MS/MS 代谢组学技术,探索汉黄芩苷发挥抗衰老作用的潜在机制。结果显示,0.02和0.5 mg/mL 汉黄芩苷均可整体延长果蝇寿命,并能够分别延长果蝇平均寿命5.64%和5.39%,延长最高寿命2.74%和5.12%;与30 d组相比,汉黄芩苷能够显著上调果蝇体内抗氧化酶基因SOD1、SOD2和CAT的表达水平,下调MTH的表达水平。果蝇代谢组学分析共找到17个潜在生物标志物,主要参与氨基酸代谢（D-谷氨酰胺和D-谷氨酸代谢,丙氨酸、天冬氨酸和谷氨酸代谢,精氨酸和脯氨酸代谢,缬氨酸、亮氨酸和异亮氨酸代谢）和能量代谢（氮代谢）。该结果表明,汉黄芩苷延缓衰老与上调抗氧化基因表达和调控不同代谢途径有关。     

Metabonomic analysis of the therapeutic effect of exendin-4 for the treatment of tBHP-induced injury in mouse glomerulus mesangial cells

Although previous studies have reported the protective effect of glucagon-like peptide-1 (GLP-1) in diabetes nephropathy, the molecular mechanism such as nephroprotection remains elusive. In this study, we explored the molecular mechanism of exendin-4 as an GLP-1 receptor agonist for the treatment of tert-butyl hydroperoxide (t-BHP)-induced injury in mouse glomerulus mesangial cells (SV40 MES 13 cells) via an NMR-based metabonomic analysis. We found that exendin-4 protected mesangial cells from t-BHP-mediated toxicity, decreased the percentage of t-BHP-treated cells undergoing apoptosis, and restored glucose consumption in the t-BHP-treated group. A supervised partial least-squares discriminant analysis (PLS-DA) revealed that the metabolic profiles could be distinguished between the control, t-BHP-treated, and exendin-4-pretreated groups. Our findings indicate that exendin-4 pretreatment can cause distinct changes in energy, glycerol phospholipid, and amino acid metabolism. Our study provides novel insight into the metabolic mechanism of exendin-4-mediated nephroprotective effects.     

Ethanol,not detectably metabolized in brain,significantly reduces brain metabolism,probably via action at specific GABA(A) receptors and has measureable metabolic effects at very low concentrations

Ethanol is a known neuromodulatory agent with reported actions at a range of neurotransmitter receptors. Here, we measured the effect of alcohol on metabolism of [3‐¹³C]pyruvate in the adult Guinea pig brain cortical tissue slice and compared the outcomes to those from a library of ligands active in the GABAergic system as well as studying the metabolic fate of [1,2‐¹³C]ethanol. Analyses of metabolic profile clusters suggest that the significant reductions in metabolism induced by ethanol (10, 30 and 60 mM) are via action at neurotransmitter receptors, particularly α4β3δ receptors, whereas very low concentrations of ethanol may produce metabolic responses owing to release of GABA via GABA transporter 1 (GAT1) and the subsequent interaction of this GABA with local α5‐ or α1‐containing GABA(A)R. There was no measureable metabolism of [1,2‐¹³C]ethanol with no significant incorporation of ¹³C from [1,2‐¹³C]ethanol into any measured metabolite above natural abundance, although there were measurable effects on total metabolite sizes similar to those seen with unlabelled ethanol.

     

代谢组研究

代谢是生命活动中所有（生物）化学变化的总称。代谢活动是生命活动的本质特征和物质基础。代谢组是生物体内源性代谢物质的动态整体。代谢组学是关于生物体内源性代谢物质的整体及其变化规律的科学。系统生物学研究的本质就是要求对研究对象的相关分子机理进行定量、普适、整体和可预测性地认识。作为全局系统生物学的基础和系统生物学的一个重要组成部分,代谢组学是以物理学基本原理为基础的分析化学、以数学计算与建模为基础的化学计量学和以生物化学为基础的生命科学等学科交叉的学科。在过去七年多的时间里,这门新兴的学科得到了迅速的发展,并已广泛地应用到了分子病理学、毒理学、功能基因组学、临床医学和环境科学等领域。本文就代谢组学的本质、代谢组分析研究方法及其应用做了概述。     

代谢组学技术及其在毒理学研究领域中的应用

代谢组学是近几年发展起来的对某一生物或细胞所有低分子量代谢产物进行定性和定量分析的一门新学科,其研究对象主要是生物体液,研究手段主要是核磁共振和质谱。简要综述了代谢组学的概念、代谢组学在毒理学研究领域中的应用、当前代谢组学研究中存在的问题及今后的发展趋势,并探讨了代谢组学在研究毒物作用机制、药物的临床前安全性评价、确定毒物作用靶器官及器官特异性新的生物标志物中的实际应用。     

虫草素产量不同的蛹虫草菌株代谢组差异

为探究虫草素高产和低产的两株蛹虫草菌代谢差异及其与虫草素代谢的关联性,本研究采用UPLC-QTOF-MS广泛靶向代谢组学技术和多元统计分析方法,结合相同蛹虫草菌不同发酵时间的菌丝体差异代谢物相对含量的变化情况,比较了两株蛹虫草菌相同发酵时间的菌丝体代谢差异.结果 表明:蛹虫草ClCC 14014菌株发酵第20和10天的...     

马钱子致大鼠体内毒性的研究*——“装袋”算法和16S rRNA基因测序技术在毒理学研究中的应用

目的 中药马钱子（Strychnos nux-vomica L.,SN）在临床上具有消肿止痛的功效,然而,由于含有生物碱类成分,马钱子具有一定毒性。人们对马钱子毒性所引起的大鼠内源性代谢变化及其对肠道微生物群代谢失调的潜在影响知之甚少,因此,马钱子的毒理学研究对其安全性评价具有重要意义。本研究将代谢组学和16S rRNA基因测序技术相结合来探索马钱子的致毒机制。方法 通过急性、蓄积性和亚急性毒性试验,分别确定马钱子的中毒剂量、毒性强度和毒性靶器官。超高效液相色谱-质谱联用技术用于分析大鼠灌胃马钱子后的血清、肝脏和肾脏样本。利用基于装袋算法的决策树和K最近邻（K nearest neighbor,KNN）模型对组学数据进行分类。从大鼠粪便中提取样本后,使用高通量测序平台对细菌的16s rRNA V3-V4区域进行分析。结果 装袋算法提高了样本分类的准确率。共鉴定出12个生物标志物,这些生物标志物的代谢失调可能是马钱子致体内毒性的原因。拟杆菌、粪厌氧棒菌、颤螺菌、双茎体菌等与肾肝功能的生理指标密切相关,这表明马钱子引起的肝肾损害可能与这些肠道细菌的代谢紊乱有关。结论 本文揭示了马钱子的体内致毒机制,为马钱子临床上的安全合理使用提供了科学依据。     

代谢组学及其应用

对代谢组学的概念、特性、发展历史做了简要介绍,综述了当前代谢组学研究中的数据采集、数据分析中采用的技术,及代谢组学在疾病诊断、药物毒性研究、植物和微生物等邻域的应用,并对代谢组学的发展作了展望。