基于代谢组学的细胞内源性代谢物研究进展

代谢组学是继基因组学、转录组学和蛋白质组学之后新兴的一种定性和定量分析复杂生物样品中所有小分子代谢物的组学方法,也是目前组学研究领域的热点之一。代谢组学主要考察生物体系受刺激或扰动后内源性代谢物的变化,是系统生物学的有机组成部分。近年来在代谢组学研究中,细胞内源性代谢物的研究已取得了很大进展。本文结合代谢组学的基本含义,综述了细胞样品前处理方法及代谢靶向分析、代谢轮廓分析、代谢组学分析和代谢足迹分析在细胞方面的研究进展。     

代谢组学技术的整合运用及其在中药现代化中的应用展望

代谢组学已经成为当今生物与医药领域的研究热点，作为系统生物学的核心组学，与其他组学进行的系统性研究为中药作用机制研究提供了思路和方法。质谱、核磁共振等现代分析技术作为代谢组学研究的技术平台，它们的整合运用加速了代谢组学的研究进程。本文就代谢组学技术的整合运用及研究现状作了简要的概述，并对代谢组学在中药现代化过程中的研究前景进行了展望。     

代谢组学研究技术进展

目的 介绍代谢组学研究技术的最新进展。方法 综合国内外文献报道,介绍当前代谢组学研究中样品制备、仪器分析技术、数据处理方法和结果分析的最新研究概况。结果 代谢组学研究技术取得了一定进步,拓宽了代谢组学的应用范围。结论 自动化、标准化、整合化和完整化将是代谢组学研究技术的发展方向。     

代谢组学研究技术进展

目的介绍代谢组学研究技术的最新进展。方法综合国内外文献报道,介绍当前代谢组学研究中样品制备、仪器分析技术、数据处理方法和结果分析的最新研究概况。结果代谢组学研究技术取得了一定进步,拓宽了代谢组学的应用范围。结论自动化、标准化、整合化和完整化将是代谢组学研究技术的发展方向。     

基于液质联用技术的代谢组学生物样本——血液及尿液的收集和处理

代谢组学是定性及定量描述内源性代谢物变化的科学，正广泛应用于疾病诊断和预后过程中。血液和尿液是代谢组学研究中常用的生物样本，但因其代谢物繁多、成分复杂，亟须使用灵敏度高、特异性强的分离分析技术。液相色谱-质谱联用（liquid chromatography-mass spectrometry， LC-MS）技术分析速度快、分离效率高，非常适用于代谢组学等方面的研究。生物样本的采集、处理过程是实验中最关键的一步，直接决定着实验结果的准确性，因此生物样本预处理过程的标准化显得尤为重要。本文综述了常用的生物流体—血液和尿液的收集、处理及储存方法，以期为有关代谢组学研究提供参考。     

代谢组学技术在临床诊断中的研究进展

代谢组学是系统生物学的重要组成部分。研究显示,代谢组学在临床诊断过程中有着不可比拟的作用。当机体受到外源性或内源性刺激时,体内小分子代谢物的种类及含量会发生显著改变,代谢组学技术通过对体内复杂代谢物的动态变化进行分析,分析代谢物变化与机体病理或生理变化的直接相关性。随着分析技术的不断发展,核磁共振、色谱-质谱联用等先进技术被广泛应用于代谢组学的研究中。目前临床运用代谢组学技术可以全面分析患者体液中代谢物的动态变化,发现体液中与疾病密切相关的特征性生物标志物,从而实现疾病的早期临床诊断。本文主要对代谢组学技术及其在临床诊断过程中的广泛应用作一综述。     

代谢组学的发展与药物研究开发

代谢组学是近年来新发展起来的一门组学,其主要研究体系有生物体液、生物组织及单个细胞的代谢组,利用一些现代的分析技术,如NMR、LC-MS、GC-MS等,取得整个研究体系的多维数据后,利用模式识别和专家系统技术寻找其中的系统生物学信息。本文从代谢组学的发展,代谢组学的研究范围和研究方法,以及在药物的作用机制和安全性评价,疾病模型,特别是中药研究的应用等方面予以阐述。     

代谢组学技术在骨质疏松症中的应用研究进展

目的 总结代谢组学技术在骨质疏松症中的应用情况。方法 检索万方、PubMed数据库自建库起至2023年5月有关代谢组学技术用于骨质疏松症的发病机制、诊断及防治的文献。结果 骨质疏松症主要由氨基酸、脂质、能量等的代谢紊乱引起,其差异代谢物均可作为疾病预测、诊断、治疗的生物标志物。代谢组学与多组学的联用是目前的研究热点。结论 代谢组学具有巨大的临床翻译潜力,从疾病预测到治疗,为进行全面的骨质疏松症研究提供了新的方向。     

代谢组学技术在神经精神性疾病中的应用研究进展

神经精神性疾病的发病过程,一般与氧化应激、神经递质和脂肪酸代谢紊乱及线粒体功能异常有关。代谢组学技术则能通过分析比较不同病理生理状态下体内小分子代谢产物,全面监测多条与疾病相关的代谢通路,进而寻找有价值的生物标志物,揭示其病理过程。本文主要对代谢组学技术在精神分裂症、抑郁症、帕金森等神经精神性疾病中的应用进行了综述。     

以抑郁症葡萄糖分解代谢障碍机制解析为例探究稳定同位素示踪代谢组学的研究思路与策略

稳定同位素示踪代谢组学通过示踪标记的原子对整个机体代谢过程进行追踪分析,属于生物医药领域前沿技术。该技术对于阐释疾病发生机制、发现疾病的生物标志物和药物作用靶点具有重要意义与价值。本文以抑郁症葡萄糖分解代谢障碍机制解析为例,系统地阐述稳定同位素示踪代谢组学技术及其应用。提出了在非标记代谢组学基础上的稳定同位素示踪代谢组学研究思路,给出了从代谢物同位素丰度、关键代谢酶、代谢物流向和代谢物流量四个维度展开生物学意义阐释的研究策略,突破了基于整体动物实验的稳定同位素示踪代谢组学研究技术瓶颈,为该技术的推广应用提供科学依据。     

Biomarkers,metabonomics, and drug development: Can inborn errors of metabolism help in understanding drug toxicity?

Application of "omics" technology during drug discovery and development is rapidly evolving. This review evaluates the current status and future role of "metabonomics" as a tool in the drug development process to reduce the safety-related attrition rates and bridge the gaps between preclinical and clinical, and clinical and market. Particularly, the review looks at the knowledge gap between the pharmaceutical industry and pediatric hospitals, where metabonomics has been successfully applied to screen and treat newborn babies with inborn errors of metabolism. An attempt has been made to relate the clinical pathology associated with inborn errors of metabolism with those of drug-induced pathology. It is proposed that extending the metabonomic biomarkers used in pediatric hospitals, as "advanced clinical chemistry" for preclinical and clinical drug development, is immediately warranted for better safety assessment of drug candidates. The latest advances in mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy should help replace the traditional approaches of laboratory clinical chemistry and move the safety evaluation of drug candidates into the new millennium.     

代谢组学的研究现状与展望

代谢组学是20世纪90年代中期发展起来的对某一生物或细胞所有低相对分子质量代谢产物进行定性和定量分析的一门新学科,由于其广泛的应用前景,目前已成为系统生物学的重要组成部分。现简要介绍了代谢组学的含义、代谢组学研究的历史沿革、当前代谢组学研究中的分析技术、数据解析方法,综述了代谢组学在药物毒理学研究、疾病诊断、植物和中药等领域的应用情况,并对当前代谢组学研究中存在的问题及发展趋势进行探讨。     

Metabonomics in toxicology: a review.

Metabonomics and its many pseudonyms (metabolomics, metabolic profiling, etc.) have exploded onto the scientific scene in the past 2 to 3 years. Nowhere has the impact been more profound than within the toxicology community. Within this community there exists a great deal of uncertainty about whether metabonomics is something to count on or just the most recent technological flash in the pan. Much of the uncertainty is due to unfamiliarity with analytical and chemometric facets of the technology and the attendant fear of any "black-box." With those fears in mind, metabonomics technology is reviewed with particular emphasis on toxicologic applications in preclinical drug development. The jargon, logistics, and applications of the technology are covered in some detail with emphasis on recent work in the field.     

代谢组学及其在药理学中的进展

随着化学检测技术和数据处理技术的发展,代谢组学成为了基因功能研究的重要手段,该文对代谢组学的基本知识和基本技术进行了介绍,对其在药理学研究中的应用作了综述和展望。     

Application of high-throughput Fourier-transform infrared spectroscopy in toxicology studies: contribution to a study on the development of an animal model for idiosyncratic toxicity

An evaluation of high-throughput Fourier-transform infrared spectroscopy (FT-IR) as a technology that could support a "metabonomics" component in toxicological studies of drug candidates is presented. The hypothesis tested in this study was that FT-IR had sufficient resolving power to discriminate between urine collected from control rat populations and rats subjected to treatment with a potent inflammatory agent, bacterial lipopolysaccharide (LPS). It was also hypothesized that co-administration of LPS with ranitidine, a drug associated with reports of idiosyncratic susceptibility, would induce hepatotoxicity in rats and that this could be detected non-invasively by an FT-IR-based metabonomics approach. The co-administration of LPS with "idiosyncratic" drugs represents an attempt to develop a predictive model of idiosyncratic toxicity and FT-IR is used herein to support characterization of this model. FT-IR spectra are high dimensional and the use of genetic programming to identify spectral sub-regions that most contribute to discrimination is demonstrated. FT-IR is rapid, reagentless, highly reproducible and inexpensive. Results from this pilot study indicate it could be extended to routine applications in toxicology and to supporting characterization of a new animal model for idiosyncratic susceptibility.     

Predictive strategy for the rapid structure elucidation of drug degradants

Structural information on drug degradants and impurities can serve to accelerate the drug discovery and development cycle. Traditional structure elucidation methodologies for obtaining this information are often slow and resource-consuming; therefore, LC/MS profiling and LC/MS/MS substructural analysis methodologies have been developed to rapidly and accurately elucidate structures of impurities and degradants. This work is a further development of methodologies used for the elucidation of degradation products of paclitaxel [K.J. Volk et al., Proc. 9th AAPS Ann. Meeting, 1994, p. 29]. In this study cefadroxil was used as a model compound for the evaluation of a predictive strategy for the production and elucidation of impurities and degradants induced by acid, base, and heat, using LC/MS and LC/MS/MS profiling methodology, resulting in an LC/MS degradant database which includes information on molecular structures, chromatographic behavior, molecular weight, UV data, and MS/MS substructural information. Furthermore, libraries such as this can provide a predictive foundation for pre-clinical development work involving drug stability, synthesis, and monitoring.     

Depsipeptide (FR901228, NSC-630176) pharmacokinetics in the rat by LC/MS/MS

Depsipeptide, a cyclic peptide (FR), isolated from Chrombacterium violaceum strain WB968 by Fujisawa Company during a screening program for anti-oncogene agents, possesses potent antitumor activity against human tumor cell lines and xenografts. This compound has been selected for preclinical and early clinical development by the National Cancer Institute. The pharmacokinetics and oral bioavailability of this depsipeptide in the rat were investigated in the present study. A sensitive and specific electrospray LC-tandem mass spectrometry method was first developed and validated for the analysis of this depsipeptide in plasma using t-boc--d-glutamic acid benzyl ester as the internal standard. The routine sensitivity limit was 1 or 10 ng/ml using 1.0 or 0.1 ml of plasma sample. The within-run CV values were 11.8, 17.9, 11.0, and 5.0% at 1, 10, 100, and 500 ng/ml levels, respectively, with corresponding accuracy of 94.4, 109, 95, and 97% (all n=6). A formulation based on ethanol, normal saline and PEG400 was then developed and Fischer rats were given this formulated drug separately by intravenous and oral route. Plasma drug concentrations were measured by this method and pharmacokinetics were analyzed by the standard techniques. Plasma concentration-time profiles were found to follow a biexponential decline with a mean terminal t1/2 of 97 min and mean total clearance (CLt) of 425.3 ml/min/kg following iv dosing at 10 mg/kg. Following oral dosing at 50 mg/kg, the peptide was absorbed but produced erratic drug levels also with a bioavailability of 15.6%. Thus, active plasma concentrations can be produced up to 3 hrs in the rat following a single dose at 10 mg/kg and the peptide represents one of the very few orally absorbed peptides reported.     

Application of metabonomics in a compound ranking study in early drug development revealing drug-induced excretion of choline into urine

Selecting drug candidates based on toxicity is an important step in early drug development. In this case study, it is shown how metabonomics is applied to a ranking study, in which drug candidates with equal pharmacological activities are selected based on least toxic side effects. The metabonomic analyses were carried out on an animal study that followed an established protocol for pilot toxicology/ranking studies in rats, however, not specifically modified for a metabonomic assessment. It is shown how conditions not specificially adopted for metabonomics investigations can significantly influence the metabolic profiles recorded by NMR. Furthermore, it is shown how the multivariate analysis of the NMR spectra identified an extreme excretion of an endogenous metabolite into urine induced by two out of the five drug candidates. The subsequent structure elucidation by two-dimensional NMR experiments and a subsequent validation by spiking experiments identified the metabolite as choline. The discussion of the mechanistic background for the excretion of choline, which is usually well-conserved in the body, results in two hypotheses of either a massive degradation of cell membranes or an inhibition of the choline oxidation. Although the validation of these hypotheses needs a follow-up study, the finding of a increased excretion of the important metabolite choline warrants exclusion of these two compounds as viable drug candidates from a metabonomics point of view.