Metabolomic Profilings of Urine and Serum from High Fat-Fed Rats via 1H NMR Spectroscopy and Pattern Recognition

¹H NMR spectroscopy in combination with multivariate statistical analysis was applied to explore the metabolic variability in urine and serum of high fat-fed rats relative to normal chow-fed ones. Metabolites contributing to intergroup discrimination identified by partial least squares discriminant analysis include 3-hydroxybutyrate, glutamate, glutamine, citrate, choline, hippurate, alanine, lactate, creatinine, taurine, acetate, etc. The aging effect along with long-term feeding was delineated with metabolic trajectory in principal component analysis score plot and age-related differences on metabolic profiling under different dietary intervention were recognised. The identified metabolites responsible for obesity were all imported into a web tool for network-based interpretation of compound lists to interpret their functional context, molecular mechanisms and disturbed signalling pathway globally and systematically. The results are useful for interpreting the pathology of obesity and further probing into the relationship between dietary-induced obesity and type 2 diabetes mellitus.     

黄芪口服液降低大鼠顺铂毒性作用机制的尿液代谢组学研究

采用基于液相色谱-飞行时间质谱联用(LC-TOF-MS)技术的代谢组学方法,分析大鼠尿液内源性代谢物的变化,研究黄芪口服液(HO)降低大鼠顺铂(CDDP)毒性的作用机制.采用低剂量多次腹腔注射CDDP的方法建立CDDP染毒大鼠模型,并连续给予16天HO.于第18天收集正常对照(Control)组、顺铂模型(CDDP)组和黄芪口服液(HO)组大鼠的24 h尿液, 进行LC-TOF-MS分析,以获取尿液代谢物组数据集,对所得数据进行主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)等多元统计分析,以筛选潜在生物标志物.于第20天采集大鼠血清测定肌酐和尿素氮水平.血清指标测定结果表明, HO可以显著降低CDDP染毒大鼠的肌酐和尿素氮水平(p<0.05).PCA得分图显示,3组可分别聚类,HO组位于Control组和CDDP组中间,表明HO可部分改善CDDP所致大鼠尿液代谢产物的异常变化.综合OPLS-DA分析、t检验和倍数变化分析结果,最终共筛选并初步鉴定出35个尿液代谢产物作为HO减毒相关的潜在生物标记物.代谢通路分析结果表明,HO可通过纠正体内氨基酸代谢、能量代谢和核苷酸代谢等通路的紊乱,降低CDDP所致机体毒性.     

Metabonomic Studies Comparing Capillary and Conventional HPLC-oa-TOF MS for the Analysis of Urine from Zucker Obese Rats

The application of capillary HPLC and orthogonal acceleration (oa) TOF-MS to a metabonomic investigation of the urinary metabolic profiles of samples obtained from male and female Zucker rats is described. The results obtained using capillary HPLC for performing the chromatographic separation are also compared with the analysis of the same samples by conventional HPLC. Capillary HPLC-MS gave increased sensitivity (ca. 100 fold) despite using a fraction of the sample volume consumed by HPLC (0.5 L vs 10 L). Capillary HPLC-MS also provided a greater peak count (ca. 3000 ions compared to 1500 for HPLC) for the same analysis time compared to conventional HPLC-MS. These data were processed using partial least squares discriminant analysis (PLS-DA)) enabling markers of diurnal variation to be detected for urine samples from both male and female rats.     

大鼠尿液中游离皮质酮昼夜节律的分析研究

为探讨大鼠尿液游离皮质酮昼夜节律性变化,采集24 h内8个时间段大鼠尿液,应用ELISA法检测了尿游离皮质酮含量,生化法检测了尿肌酐含量,以皮质酮与尿肌酐比值反映尿游离皮质酮水平,对各个时间段尿游离皮质酮水平的生物时间序列进行了宏观分析与比较.结果表明,大鼠尿液游离皮质酮与尿肌酐比值呈现昼夜波动性;尿液游离皮质酮的检测...     

Biochemical and Molecular Investigation of the Effect of Saponins and Terpenoids Derived from Leaves of Ilex aquifolium on Lipid Metabolism of Obese Zucker Rats

Ilex paraguariensis, the holly tree, is a plant with recognized biological properties, whose aqueous infusions are known as “Yerba mate”, that regulate lipid metabolism, reduce obesity, and improve brain stimulation. In the present study, the effect of standardized saponin and terpenoid fractions of a European taxon, Ilex aquifolium, on blood biochemical parameters in a rat model of metabolic disorder, (fa/fa) Zucker, are presented. The profiles of the volatile fractions of two species and six European varieties of Ilex were investigated. After selecting the best variety, the saponin and terpenoid fractions were isolated and standardized, and animals were fed 10 mg kg⁻¹ b.w. for 8 weeks. A statistically significant decrease in liver adiposity was observed, confirmed by histology and quantitative identification (gas chromatography–mass spectrometry analyses of hepatic lipids. RT-qPCR analysis of gene expression in the aorta revealed that the administration of the terpenoid fraction downregulated LOX-1, suggesting a reduction in atherosclerotic stimuli. In addition, a statistically significant reduction (p < 0.05) in PPARγ for the saponin fraction was observed in the liver. The expression of the ACAT-1 gene in the liver, responsible for the formation of cholesterol esters, increased significantly in the group receiving the terpenoid fraction compared to the control, which was also confirmed by the analysis of individual blood biochemical parameters. The opposite effect was observed for saponins. Taking the above into account, it is shown for the first time that Ilex aquifolium can be a source of compounds that positively influence lipid metabolism.     

银杏叶提取物对大鼠抗氧化能力的影响

探讨了银杏叶提取物对大鼠抗氧化能力的影响.结果表明:银杏叶提取物可以增加运动机体抗氧化酶的活性,提高机体的抗氧化功能,减轻大强度耐力训练对大鼠组织造成的脂质过氧化损伤,改善运动造成的自由基代谢紊乱,加快自由基的清除.     

The Effect of Blood Contained in the Samples on the Metabolomic Profile of Mouse Brain Tissue: A Study by NMR Spectroscopy

(1) Recently, metabolic profiling of the tissue in the native state or extracts of its metabolites has become increasingly important in the field of metabolomics. An important factor, in this case, is the presence of blood in a tissue sample, which can potentially lead to a change in the concentration of tissue metabolites and, as a result, distortion of experimental data and their interpretation. (2) In this paper, the metabolomic profiling based on NMR spectroscopy was performed to determine the effect of blood contained in the studied samples of brain tissue on their metabolomic profile. We used 13 male laboratory CD-1^® IGS mice for this study. The animals were divided into two groups. The first group of animals (n = 7) was subjected to the perfusion procedure, and the second group of animals (n = 6) was not perfused. The brain tissues of the animals were homogenized, and the metabolite fraction was extracted with a water/methanol/chloroform solution. Samples were studied by high-frequency ¹H-NMR spectroscopy with subsequent statistical data analysis. The group comparison was performed with the use of the Student’s test. We identified 36 metabolites in the brain tissue with the use of NMR spectroscopy. (3) For the major set of studied metabolites, no significant differences were found in the brain tissue metabolite concentrations in the native state and after the blood removal procedure. (4) Thus, it was shown that the presence of blood does not have a significant effect on the metabolomic profile of the brain in animals without pathologies.     

亚硒酸钠对大鼠代谢产物影响的核磁共振研究

本文采用核磁共振(^1H NMR)技术研究了在最高耐受摄入量条件下补充亚硒酸钠对大鼠尿液代谢产物的影响。结果表明,在大鼠尿液中甲酸、乙酸、乳酸、丙氨酸、琥珀酸、甘氨酸、马尿酸、苯丙氨酸、色氨酸等代谢物谱峰信号显著增强,而柠檬酸、肌酸、尿素、尿甘素和氧化三甲胺(TAMO)的谱峰信号则显著降低。乳酸、柠檬酸和琥珀酸是体内三羧酸循环的中间产物,其代谢异常是能量代谢紊乱的标志;马尿酸、苯丙氨酸、丙氨酸的代谢异常与肾小球的滤过和回收功能有关;乙酸和甘氨酸的代谢异常则是肝功能损伤的标志,组织切片的结果也证明大鼠的肝和肾已经发生损伤,与^1H NMR的结果相一致。采用HPLC检测到补充亚硒酸钠后尿液中8-OHdG(8-hydroxy1-2′-deoxyguansoine)水平显著升高,证明体内氧化损伤的发生。以上结果表明,在最高耐受摄入量条件下补充亚硒酸钠是不安全的,用NMR方法对阐明硒的毒理学分子基础是有效的。     

硒对氟中毒大鼠尿氟及脑肾组织脂质过氧化物含量的影响

大鼠饲料分别加入２×１０－６、４×１０－６硒．饮用７．５×１０－６、１５×１０－６氟水．１个月后，随尿硒粪硒排量增加，尿氟排量也增高，以２×１０－６硒饲料排氟效果最佳．血浆中ＡＬＰ、ＧＯＴ、ＧＰＴ及ＬＤＨ等酶活性加硒组有不同程度的降低．随后两次增高氟水量重复试验，给以２×１０－６硒饲料，仍观察到尿氟排量增高．血中含硒酶谷胱甘肽过氧化物酶活性也有升高趋势，脑及肾中脂质过氧化物（ＬＰＯ）含量明显减少，病区患者服硒后，尿氟含量亦高于对照组．实验结果提示；１）硒能促使体内氟由尿中排出；２）降低某些组织ＬＰＯ含量；３）减少体内氟含量；４）预防和改善氟中毒某些临床症状。     

Inhibitory Effect of Thai Purple Rice Husk Extract on Chemically Induced Carcinogenesis in Rats

This study investigated the cancer chemopreventive effects of an acidic methanol extract of purple rice husk on chemically induced carcinogenesis in rats. This purple rice husk extract (PRHE) had high polyphenol contents. Vanillic acid was a major phenolic compound in PRHE. Three major anthocyanins found in PRHE were malvidin-3-glucoside, peonidin-3-glucoside and cyanidin-3-glucoside. PRHE was not toxic and clastogenic in rats. The LD₅₀ of PRHE was greater than 2000 mg kg⁻¹ body weight (BW). The oral administration of 300 or 1000 mg kg⁻¹ BW of PRHE for 28 days significantly decreased the number of micronucleated hepatocytes in diethylnitrosamine-initiated rats. The inhibitory mechanisms were associated with the reduction of cytochrome P450 2E1 expression and induction of some detoxifying enzymes in the liver. In addition, treatment with 500 mg kg⁻¹ BW of PRHE for eight weeks did not induce preneoplastic lesions in the liver and colon. It significantly inhibited hepatic glutathione-S-transferase positive foci formation induced by diethylnitrosamine and 1,2-dimethylhydrazine by suppression of hepatocyte proliferation and induction of apoptosis. In conclusion, PRHE did not present toxicity, clastogenicity or carcinogenicity in rats. It exhibited cancer chemopreventive properties against chemically induced early stages rat hepatocarcinogenesis. Anthocyanins and vanillic acid might be candidate anticarcinogenic compounds in purple rice husk.     

生物酶振荡器研究谷胱甘肽的抗氧化效应

组装了一种生物酶振荡器（Peroxidase-Oxidase Biochemical Oscillator） 的实验装置、建立了相应的实验方法，在此基础上得到单倍周期、四倍周期、混沌 等丰富的振荡图谱信息，并采用它作为“探针”，检测了谷胱甘肽（GSH）的抗氧 化效应，它可作为定量分析的基础，线性范围为9.38 * 10~(-7) ～ 7.50 * 10~(- 5) mol·L~(-1)，探讨了某些可能的反应机理。     

生物酶振荡器研究谷胱甘肽的抗氧化效应

组装了一种生物酶振荡器（Peroxidase-Oxidase Biochemical Oscillator） 的实验装置、建立了相应的实验方法,在此基础上得到单倍周期、四倍周期、混沌 等丰富的振荡图谱信息,并采用它作为“探针”,检测了谷胱甘肽（GSH）的抗氧 化效应,它可作为定量分析的基础,线性范围为9.38 * 10~(-7) ～ 7.50 * 10~(- 5) mol·L~(-1),探讨了某些可能的反应机理。     

Effect of Nanofluid Treatment on Water Sensitive Formation to Investigate Water Shock Phenomenon,An Experimental Study

Permeability reduction in porous media as a result of frail and tenuous fine particles migration would decrease the productivity index in the subterranean reservoirs. During reservoir stimulation by injecting fluids into the reservoir, as the salinity condition of the formation brine changes, fine particles initiate the triggering process. In this study, MgO-based nanofluid as a fines fixation agent was stably prepared based on the particle size distribution and characterized through transmission electron microscopy analysis. Afterward, several core flooding tests were performed using Berea sandstone cores to study the effect of nanofluid injection on fines fixation in the water shock phenomenon. Permeability reduction occurred up to 95% of inchoate permeability for no treatment case, which was also confirmed by scanning electron microscopy analysis. Finally, MgO nanofluid with 0.03 wt% concentration and 120 minutes soaking time in the core mitigated the fine particles release and fixed them on the pore walls' surfaces critically reducing the formation damage. The analysis shows that outweighing the attraction potentials compared to repulsions was the main mechanism after nanofluids treatment.     

核磁共振碳谱中的重原子效应

介绍了核磁共振碳谱中的重原子效应,分析了溴和碘的重原子效应对不同杂化状态和环境下碳的化学位移的影响,并总结了其影响规律。     

稀土对大鼠尿液成分影响的核磁共振研究

目前,稀土生物效应的研究已进入了分子和细胞水平,但在活体动物内研究稀土的作用情况还不多见。稀土化合物随食物进入动物体内后,会对动物体内的器官、组织、细胞产生一定影响,将导致其体液容量、分布、电解质浓度等方面的变化。现代NMR技术是目前药理毒理学研究的有效手段之一,它具有简便、全面、快捷、非破坏性等优点,可用于对体液样品的检测和研究。本文采用现代核磁共振技术研究了稀土化合物硝酸镧灌胃给药后对Wistar大鼠尿液中代谢产物的影响,为阐明稀土化合物的毒理学机制和稀土的进一步开发利用提供理论依据。     

骨髓增生异常综合症患者血液红细胞^31P—NMR谱的特征

本文介绍了用３１Ｐ－ＮＭＲ谱研究骨髓增生异常综合症（ＭＤＳ）患者血液红细胞的方法。分析了患者和正常人的血液红细胞在３１Ｐ－ＮＭＲ谱上的差异，探讨这些差异在临床医学上对ＭＤＳ患者进行诊断和疗效跟踪检测的应用途径。     

Antibacterial and Antiproliferative Activities of Azadirachta indica Leaf Extract and Its Effect on Oil-in-Water Food Emulsion Stability

The present study aims to identify and quantify the phenolic compounds of Azadirachta indica leaf extract using HPLC-MS and to evaluate the antioxidant, antibacterial (against different Gram-positive and negative bacteria) and in vitro anti-proliferative activities of this extract (against breast, human liver and cervix adenocarcinoma-derived cells). The application of this extract as a natural antioxidant for food preservation was also tested on oil-in-water food emulsions for the first time in the present work in order to determine the use of Azadirachta indica leaves as a natural additive to preserve the food against lipid oxidation and rancidity. The results obtained revealed that 50%-aqueous ethanol leaf extract showed the best extraction yield (25.14%), which was characterized by a high content in phenolic compounds and strong antioxidant activity. Moreover, this leaf extract inhibited the growth of the bacterial strains tested (Staphylococcus aureus, Escherichia coli, Salmonella paratyphi and Micrococcus luteus) and showed better anti-proliferative activity against breast and cervix adenocarcinoma-derived cells than human liver cancer cells after 48 h of treatment. Additionally, Azadirachta indica leaf extract showed almost similar effects as gallic acid solutions (0.25% and 0.5%) in preserving the oxidation of oil-in-water food emulsions and prevented the formation of secondary oxidation products (malondialdehyde) as well. The results obtained suggested that extracts of Azadirachta indica leaves are a potential source of antioxidant and antibacterial compounds and pointed to the potential of these natural extracts as therapeutic agents.     

NMR Studies on the Subacute Biochemical Effects of Aristolochic Acid on Rat Serum

The subacute effect of aristolochic acid （AA） on rat serum was studied by NMR method. The biochemical effects induced by AA were characterized by an increase in the amounts of creatinine, trimethylamine N-oxide, acetoacetate, acetate and 3-D-hydroxybutyrate and lactate in serum from ^1H NMR spectra. Principal component analysis was used for further comparing the similarities of ^1H NMR spectral profiles of serum from rats treated with AA and model toxins.     

硒抗大鼠肺纤维化的实验研究

为了解硒对肺纤维化的预防作用，采用油酸大鼠尾静脉注射制作肺纤维化模型，观察了硒对损伤肺组织中中性粒细胞聚集及氧自由基的干预作用。结果显示，硒可通过抑制中性粒细胞而减少氧自由基的产生，进而控制肺泡炎变、渗出，减轻炎性因子对肺脏的慢性刺激，预防肺纤维化，因此硒对油酸所致的肺纤维化有一定的预防作用。