Metabolic profiling of bile in cholangiocarcinoma using in vitro magnetic resonance spectroscopy

Objectives:

Cholangiocarcinoma (CCA) has a poor prognosis and its aetiology is inadequately understood. Magnetic resonance spectroscopy (MRS) of bile may provide insights into the pathogenesis of CCA and help identify novel diagnostic biomarkers. The aim of this study was to compare the chemical composition of bile from patients with CCA with that of bile from patients with benign biliary disease.

Methods:

Magnetic resonance spectra were acquired from the bile of five CCA patients and compared with MRS of control bile from patients with benign biliary disease (seven with gallstones, eight with sphincter of Oddi dysfunction [SOD], five with primary sclerosing cholangitis [PSC]). Metabolic profiles were compared using both univariate and multivariate pattern-recognition analysis.

Results:

Univariate analysis showed that levels of glycine-conjugated bile acids were significantly increased in patients with CCA, compared with the benign disease groups (P= 0.002). 7β primary bile acids were significantly increased (P= 0.030) and biliary phosphatidylcholine (PtC) levels were reduced (P= 0.010) in bile from patients with CCA compared with bile from gallstone patients. These compounds were also of primary importance in the multivariate analysis: the cohorts were differentiated by partial least squares discriminant analysis (PLS-DA).

Conclusions:

These preliminary data suggest that altered bile acid and PtC metabolism play an important role in CCA aetiopathogenesis and that specific metabolites may have potential as future biomarkers.     

决明子调脂作用的代谢组学研究

目的  运用代谢组学方法考察决明子水提物对高脂血症大鼠血液内源性代谢物的影响。方法  建立高脂血症大鼠模型,采用决明子水提物灌胃治疗,利用主成分分析法和偏最小二乘-判别分析对空白对照组、模型组及治疗组大鼠血液进行代谢组学分析。观察3组代谢轮廓差异并寻找生物标志物。结果  利用主成分分析法和偏最小二乘-判别分析能将样本分开,谱库分析得到9种生物标志物：L-缬氨酸、丙氨酸、甘氨酸、富马酸、异亮氨酸、丝氨酸、硬脂酸、亚油酸和胆固醇。结论  基于气相色谱-质谱联用仪的代谢组学可以很好的从内源性代谢物的变化过程反应决明子水提物对大鼠的调脂作用。

     

血浆代谢组学研究用于肝移植物功能变化的评估

目的探讨基于核磁共振波谱技术的代谢组学方法在肝移植患者围手术期肝功能评价中的应用。方法肝移植患者15例,分别于术前1d、术后1d及1周,空腹收集其血液,对丙氨酸氨基转移酶、天冬氨酸氨基转移酶、碱性磷酸酶、白蛋白、总胆红素、葡萄糖、甘油三酯、胆固醇以及乳酸等血浆生化指标进行检查,同时测定其核磁共振氢谱,进行代谢组学分析。结果代谢组学分析显示,与术前相比,术后1d患者血浆中的血糖和乳酸水平显著升高,缬氨酸、丙氨酸、丙酮、琥珀酸、谷氨酰胺、胆碱含量亦显著升高;至术后1周,血脂和胆碱水平显著升高,血糖和氨基酸类代谢物含量显著降低。主成分分析显示,肝移植围手术期各时段血样的代谢谱图之间差异显著,并能够互相区分开来。造成组间差异的主要影响因素是血糖、血脂、胆碱、乳酸等物质含量的变化。其中血糖、血脂(包括甘油三酯和胆固醇)以及乳酸的变化规律与临床检测血浆生化的改变一致,但血液中胆碱的含量随着治疗进程出现显著的变化。结论代谢组学分析方法能较清晰地反映肝移植患者围手术期各时间段血浆样本间的代谢差异和变化,作为一种辅助手段,其在肝移植患者术后诊断中可能有良好的研究和应用前景。     

肺癌痰湿蕴肺证、气阴两虚证患者呼出气冷凝液的代谢组学研究

目的:通过分析肺癌痰湿蕴肺证和气阴两虚证患者的呼出气冷凝液(EBC)代谢产物,探讨肺癌中医证型科学客观化的可行性。方法:选择肺癌痰湿蕴肺证患者(痰湿蕴肺组)21例、气阴两虚证患者(气阴两虚组)21例及健康志愿者(健康对照组)20例,采集受试者EBC,利用气相色谱-质谱(GC-MS)技术分析EBC代谢产物。结果:痰湿蕴肺组和气阴两虚组均能够与健康对照组区分,发现9种潜在的肿瘤标志物,其中3种为痰湿蕴肺组和气阴两虚组特有化合物,6种为痰湿蕴肺组和气阴两虚组与对照组共有化合物;痰湿蕴肺组能够与气阴两虚组区分,3-氨基-2-苯甲酰基-4,5,6,7-四氢苯并[b]噻吩为痰湿蕴肺组特有化合物,1-(苯基磺酰基)吡咯为气阴两虚组特有化合物。结论:EBC的代谢组学研究方法具有用于肺癌的诊断及肺癌不同中医证型的临床客观化指标的研究的可能性。     

Metabolic characterization of distinct neuroanatomical regions in rats by magic angle spinning 1H nuclear magnetic resonance spectroscopy.

High-resolution magic angle spinning (HRMAS) (1)H NMR spectroscopy has been applied to the biochemical characterization of specific brain regions in rats in order to establish baseline levels of tissue metabolite profiles with which to compare models of neuropathology or toxic lesion. Cores of tissue (20 mg) from the brain stem, cerebellum, frontal cortex, and hippocampus were obtained from histologically defined coronal slices of brain from 18 male Sprague-Dawley rats. HRMAS (1)H NMR spectra were acquired for each of the regions sampled and the degree of intersample variability, as assessed by principal components analysis and discriminant analysis by projection to latent structure was found to be low. Clear region-specific differences in the biochemical profiles were observed using both comparison of metabolite ratios and/or pattern recognition methods. Relatively low concentrations of GABA in the cerebellum, high concentrations of taurine and N-acetylaspartate in the cortex, and high levels of choline, glycerophosphocholine, and phosphocholine in the hippocampus predominantly influenced the classification of the different brain regions. Additionally, N-acetylaspartylglutamate was detected in the brain stem, but was largely absent from the other regions examined. Such analyses provide a baseline reference for further HRMAS NMR spectroscopic studies to monitor disease and pharmacological insults in specific regions of the brain.     

Prospective evaluation of potential toxicity of repeated doses of Thymus vulgaris L. extracts in rats by means of clinical chemistry,histopathology and NMR‐based metabonomic approach

In the field of natural extracts, research generally focuses on the study of their biological activities for food, cosmetic, or pharmacological purposes. The evaluation of their adverse effects is often overlooked. In this study, the extracts of Thymus vulgaris L. were obtained by two different extraction methods. Intraperitoneal injections of both extracts were given daily for four days to male Wistar Han rats, at two different doses for each extract. The evaluation of the potential toxic effects included histopathological examination of liver, kidney, and lung tissues, as well as serum biochemistry of liver and kidney parameters, and ¹H‐NMR‐based metabonomic profiles of urine. The results showed that no histopathological changes were observed in the liver and kidney in rats treated with both extracts of thyme. Serum biochemical investigations revealed significant increases in blood urea nitrogen, creatinine, and uric acid in animals treated with polyphenolic extract at both doses. In these latter groups, metabonomic analysis revealed alterations in a number of urine metabolites involved in the energy metabolism in liver mitochondria. Indeed, the results showed alterations of glycolysis, Krebs cycle, and β‐oxidative pathways as evidenced by increases in lactate and ketone bodies, and decreases in citrate, α‐ketoglutarate, creatinine, hippurate, dimethylglycine, and dimethyalanine. In conclusion, this work showed that i.p. injection of repeated doses of thyme extracts causes some disturbances of intermediary metabolism in rats. The metabonomic study revealed interesting data which could be further used to determine the cellular pathways affected by such treatments. Copyright © 2014 John Wiley & Sons, Ltd.