基于气相色谱-飞行时间质谱技术的佐剂性关节炎大鼠尿液代谢组学的研究

采用弗氏完全佐剂(FCA)诱导佐剂性关节炎(AA)大鼠模型,观察大鼠足趾肿胀度和踝关节组织的病理学形态变化。应用气相色谱-飞行时间质谱(GC-TOF MS)技术检测AA大鼠尿液代谢物谱,并对数据进行主成分分析(PCA)、偏最小二乘法-判别分析(PLS-DA)及正交偏最小二乘法-判别分析(OPLS-DA),探讨可能的发病机制。通过变量重要性投影值(VIP>1)和P值(<0.05),筛选出尿液中的差异代谢物。在模型组大鼠的尿液中共发现异柠檬酸、α-酮戊二酸、柠康酸、肌酸、3-羟基丁酸等20种差异代谢物。推断AA代谢组学的发病机制可能与能量代谢、氨基酸代谢、脂肪酸代谢途径有关。     

非靶向代谢组学用于藜芦妨害人参治疗脾气虚的机制研究

采用超高效液相色谱串联四级杆飞行时间质谱( UPLC/Q-TOF-MS)联用技术,通过非靶向代谢组学方法分析大鼠尿液内源性代谢物的变化,研究藜芦妨害人参发挥药效作用的机制。建立脾气虚大鼠模型,连续给药15天,测定力竭游泳时间及血液中白细胞、红细胞、血红蛋白的含量。结果表明,人参可显著提高脾气虚模型大鼠的力竭游泳时间(p﹤0.01),升高白细胞、红细胞及血红蛋白含量(p﹤0.05,p﹤0.01),藜芦对脾气虚模型大鼠各项指标无明显影响(p>0.05),人参与藜芦配伍后对脾气虚模型大鼠各项指标均无显著影响(p>0.05),表明藜芦妨害了人参发挥药效作用。采用UPLC/Q-TOF-MS技术及非靶向代谢组学的方法分析了空白组、模型组、人参组、藜芦组、参藜组对脾气虚模型大鼠的尿液代谢组差异,其中主成分分析( PCA)得分图显示各组代谢轮廓有显著差别,并通过正交偏最小二乘法-判别分析( OPLS-DA)及数据库检索,鉴定出15种人参干预调节脾气虚模型大鼠的潜在生物标志物,从中找出了7种人参藜芦配伍后减弱人参上述干预作用的潜在生物标志物,并对其涉及的代谢通路进行了系统分析。上述研究结果表明,人参藜芦配伍后妨害了人参对脾气虚模型大鼠的治疗作用,其机理可能是影响人参对体内能量代谢、免疫平衡及氧化还原反应等相关代谢的调节。     

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.     

核磁共振技术测试不同性别大鼠尿液代谢物的研究

采用核磁共振波谱技术测试不同性别大鼠尿液代谢物,分析性别因素对大鼠尿液代谢成分的影响.大鼠尿液核磁共振氢谱（1HNMR谱）结果采用主成分分析（principal component analysis,PCA）和正交偏最小二乘判别分析（orthogonal to partial least squares discriminant analysis,OPLS-DA）方法分析,得到不同性别大鼠尿液中的差异性代谢物.PCA分析结果显示2组尿液代谢成分有明显的差异,进一步进行OPLS-DA分析可以判别出2组尿液中具有差异性的代谢物.结果显示,雌性大鼠尿液中的丙氨酸、缬氨酸、鸟氨酸等氨基酸类以及乙酸、硫胺、氨基马尿酸、苯乙胺、氧氨嘧啶等代谢物含量高于雄性大鼠,差异有统计学意义（p〈0.05）.雄性大鼠尿液中的甲胺、二甲胺、三甲胺、肌酸酐、尿囊素、延胡索酸、甲酸等代谢物则明显高于雌性大鼠,差异有统计学意义（p〈0.05）.性别因素对大鼠尿液中的代谢成分有一定的影响.     

硫酸氧钒毒性的核磁共振代谢组学方法研究

采用基于核磁共振(NMR)的代谢组学方法,结合生化指标分析及组织病理学检测,研究了具有类胰岛素活性的硫酸氧钒(VOSO4)对Wistar大鼠的毒性作用.通过不同剂量的VOSO4对Wistar大鼠连续灌胃给药16d,收集大鼠的血清和尿液,并采集样品的1H NMR谱进行多变量数据统计分析来辨识其特征代谢物,然后采用TICL(a web Tool for automatic Interpretation of Compound List)方法建立特征代谢物的代谢网络模型,分析受影响的主要代谢途径及其相互关系.研究结果表明:高剂量组(45mg/kg)和低剂量组(15mg/kg)的特征代谢物含量与对照组存在明显的差异;与对照组相比,高剂量和低剂量组血清中乳酸、肌氨酸酐以及牛磺酸等代谢物的含量增加,尿液中氧化三甲胺(TMAO)、肌酐、牛磺酸和甘氨酸等代谢物的含量增加,并呈现显著的剂量依赖关系;给药组中乙酸和琥珀酸的含量都降低.这些结果说明VOSO4可能影响大鼠体内的糖代谢、脂类代谢及肠道菌群代谢等多个代谢系统,高剂量的VOSO4会导致肝脏毒性和肾脏损伤.     

基于液相色谱/电喷雾飞行时间质谱技术的结石症尿液代谢组学探讨

探索了一种简便的尿液样品前处理流程,通过反相液相色谱(RPLC)与亲水相互作用色谱(HILIC)相结合的高效液相色谱/电喷雾飞行时间质谱的联用技术(HPLC/ESI-TOFMS),建立了泌尿系结石症的尿液代谢组学模型,研究结石症患者尿液样品中的代谢物浓度水平与健康人对照组间的差异情况。采用主成分分析(PCA)及偏最小二乘法-判别分析(PLS-DA)等多变量数据分析方法,寻找结石症患者组和对照组之间的差异性代谢谱。模型结果显示:结石症患者尿液代谢物水平与健康对照组具有明显差异,寻找其中潜在的差异性标志物群组,有望将该模型用于泌尿系结石症的快速预测和诊断。     

基于气相色谱-质谱联用的血府逐瘀汤治疗大鼠颅脑损伤的血浆代谢组学研究

该文研究了血府逐瘀汤对颅脑损伤大鼠血浆代谢组的影响,采用气相色谱-质谱联用(GC-MS)技术对治疗组、模型组以及假手术组的血浆代谢物进行检测,定性定量分析了43种重要的代谢物。模型组和假手术组的t检验(t-test)结果显示13种代谢物存在显著差异。正交偏最小二乘-判别分析(OPLS-DA)分析结果也显示模型组和假手术组代谢差异明显。结合变量的投影重要性指标(VIP)及t检验结果,筛选出乳酸、组氨酸、棕榈酸、色氨酸、亚油酸、油酸、硬脂酸作为生物标志物。观测7种标志物在治疗组中第1、3、7、14 d的变化情况。结果显示:治疗组中乳酸、组氨酸、棕榈酸、亚油酸、硬脂酸的相对浓度逐渐降低,色氨酸的相对浓度先降低后升高,油酸的相对浓度先升高后降低,且7种代谢物的相对浓度在第14 d时均接近于假手术组的水平。表明血府逐瘀汤对颅脑损伤具有一定的治疗作用,乳酸、组氨酸、棕榈酸、色氨酸、亚油酸、油酸、硬脂酸7种代谢物可以作为生物标志物监测颅脑损伤的治疗效果及恢复情况。     

基于液相色谱-质谱联用技术的代谢组学方法研究薄荷烟对大鼠代谢的影响

将基于液相色谱-质谱联用(LC-MS)技术的代谢组学分析平台用于薄荷烟对大鼠代谢影响的研究。分析了3组大鼠的尿样,包括对照大鼠、吸食普通烟大鼠和吸食薄荷烟大鼠,并采用主成分分析(PCA)方法对数据进行模式识别。PCA得分图表明吸食薄荷烟大鼠与对照组大鼠尿样的代谢差异要小于吸食普通烟大鼠。从PCA载荷图中找到并鉴定了犬尿喹啉酸等8种重要代谢物。通过考察代谢物在对照大鼠、吸食薄荷烟大鼠和吸食普通烟大鼠尿样中的相对含量变化,进一步说明了烟草中添加薄荷醇可减少烟草对大鼠代谢的影响。     

基于液相色谱-质谱联用技术的心肌缺血大鼠血清和心肌组织代谢组学

采用异丙肾上腺素诱导心肌缺血大鼠模型,使用液相色谱-质谱法检测血清和心肌中的内源性成分,应用软件对已鉴定的40余种目标成分进行靶向提取,用主成分分析（PCA）、有监督偏最小二乘法判别分析（PLS-DA）对代谢组学数据进行多维度统计分析,筛选潜在生物标志物。与对照组相比,在心肌缺血模型组大鼠血清、组织中检测出18个差异代谢物,涉及精氨酸和脯氨酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、谷氨酰胺和谷氨酸代谢、牛磺酸和亚牛磺酸代谢等多条代谢通路。代谢产物可作为心肌缺血研究中的重要标志物,该研究结果有助于揭示心肌缺血的发病机制,可为临床疾病诊断提供思路。     

基于超高效液相色谱-四极杆飞行时间质谱联用技术的血瘀模型大鼠血浆代谢组学分析

采用盐酸肾上腺素加冰水浴建立急性血瘀大鼠模型，使用超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF/MS）检测空白对照组与血瘀模型组中血浆代谢物，用主成分分析（PCA）、有监督偏最小二乘法判别分析（PLS-DA）及正交偏最小二乘法判别分析（OPLS-DA）对代谢组学数据进行多维统计分析，筛选潜在生物标志物。与对照组相比，在血瘀模型组大鼠血浆中检测出46个差异代谢物，血瘀模型组中乙酰胆碱、N6，N6，N6-三甲基-L-赖氨酸、胞嘧啶、乙酰肉碱等21个代谢物显著上调，吲哚丙酸、LysoPC（14：0）等25个代谢物显著下调，可能与脂质代谢、半乳糖代谢、亚油酸代谢、不饱和脂肪酸生物合成、糖酵解、花生四烯酸代谢等通路有关。代谢产物可作为血瘀证研究中的重要标记物，该研究结果有助于揭示血瘀证的发病机制，可为临床血瘀疾病的诊断及选用药物治疗提供思路，为后续治疗手段提供参考依据。     

植物类中药中微量元素的因子分析和聚类分析

尝试利用化学计量学方法探讨微量元素含量与中药药性的相关性。对１０５味植物类中药４２种微量元素测定数据用因子分析和聚类分析进行了多因素分析。因子分析证实了一个１０因子模型合理解释这些微量元素间的相关关系；样本聚类分析证明了１０５株中药合理地聚类成不同组；     

Protective effects of Ocimum sanctum on lipid peroxidation and antioxidant status in streptozocin-induced diabetic rats

The antioxidant effects of Ocimum sanctum in experimental streptozocin-induced diabetic rats was evaluated in this study. Streptozocin, 55?mg?kg(-1) body weight, was injected intraperitoneally once daily for 30 days to induce diabetes mellitus in rats. Streptozocin-induced diabetic rats were orally treated with an aqueous extract of O. sanctum once daily for 30 days. After the experimental period, thiobarbituric acid reacting substances (TBARS) and antioxidant enzymes such as catalase, superoxide dismutase (SOD) and glutathione peroxidase were measured. Administration of O. sanctum to streptozocin-induced diabetic rats for 30 days significantly reduced the plasma level of TBARS and improved the status of the antioxidant enzymes catalase, SOD and glutathione peroxidase in vital organs such as the liver and kidney. These results confirmed that the Indian medicinal plant O. sanctum has a protective effect and it may be useful in controlling complications resulting from diabetes.     

High-performance liquid chromatographic method for the determination and pharmacokinetic study of mangiferin in plasma of rats having taken the traditional Chinese medicinal preparation Zi-Shen pill

A high-performance liquid chromatographic method for the determination and pharmacokinetic study of mangiferin in the plasma of rats that have been orally administered the traditional Chinese medicinal preparation Zi-Shen pill is established. Plasma samples taken from rats are pretreated by protein precipitation with acetonitrile. Separation of the main effective constituent mangiferin is accomplished on a C18 stationary phase and a mobile phase of methanol&-water (25:75, v/v) with 0.6% glacial acetic acid. The UV detection wavelength is set at 320 nm, and the detection limit for mangiferin in plasma is 0.163 micro g/mL. After validation, the method is used to take a limited view of pharmacokinetic profiles of the traditional Chinese medicinal preparation Zi-Shen pill.     

Identification of the chemical constituents of Chinese medicine Yi‐Xin‐Shu capsule by molecular feature orientated precursor ion selection and tandem mass spectrometry structure elucidation

The incomplete identification of the chemical components of traditional Chinese medicinal formula has been one of the bottlenecks in the modernization of traditional Chinese medicine. Tandem mass spectrometry has been widely used for the identification of chemical substances. Current automatic tandem mass spectrometry acquisition, where precursor ions were selected according to their signal intensity, encounters a drawback in chemical substances identification when samples contain many overlapping signals. Compounds in minor or trace amounts could not be identified because most tandem mass spectrometry information was lost. Herein, a molecular feature orientated precursor ion selection and tandem mass spectrometry structure elucidation method for complex Chinese medicine chemical constituent analysis was developed. The precursor ions were selected according to their two‐dimensional characteristics of retention times and mass‐to‐charge ratio ranges from herbal compounds, so that all precursor ions from herbal compounds were included and more minor chemical constituents in Chinese medicine were identified. Compared to the conventional automatic tandem mass spectrometry setups, the approach is novel and can overcome the drawback for chemical substances identification. As an example, 276 compounds from the Chinese Medicine of Yi‐Xin‐Shu capsule were identified.     

火焰原子吸收光谱法测定红松松针中微量元素含量

采用火焰原子吸收光谱法测定了长白山红松松针中K、Ca、Mg、Zn、Cu、Fe和Mn 7种金属元素含量。结果表明,在常量元素中K、Ca含量较高,微量元素中Fe、Zn含量较高,7种金属元素含量由高到低顺序分别为:K、Ca、Fe、Mg、Zn、Mn、Cu。可见松针中含有丰富的与健康密切相关的微量元素,具有较高的食用和药用价值。     

基于人工智能-多元多息指纹图谱探索中药一法通识品种鉴定新方法

该文建立了一种基于中药多元多息指纹图谱联合人工智能识别的中药一法通识品种鉴定新方法,方法先通过对同一味药材采用不同处理手段得到具有不同性质的化学成分信息,并构建一种集反相色谱法、亲水色谱法以及分子排阻色谱法为一体的具有普适性的多元多息指纹图谱采集方式,实现了药材中小极性小分子、大极性小分子及大分子类化合物的全面表征,再对采集的多元多息指纹图谱进行数据标准化处理,采用卷积神经网络识别不同品种中药材,获得了准确率达92%的识别模型。该方法能够对中药品种进行快速、准确、高效地鉴定,克服了传统中药品种鉴别中的主观色彩,能够更加客观精准地给出鉴定结果。     

超高效液相色谱-质谱法同时测定舒血宁注射剂中20种黄酮醇苷与萜类内酯成分

建立了舒血宁注射剂中16种黄酮及4种萜类内酯同时定量的超高效液相色谱-质谱分析方法,并采用所建立的方法同时测定了不同厂家舒血宁注射剂产品中上述20种化合物的含量。注射剂样品经甲醇-水(体积比1∶1)稀释后,在Acquity UPLC BEH Shield RP18(2.1 mm×100 mm,1.7μm)色谱柱上分析,以0.1%(体积分数)甲酸水溶液-乙腈为流动相进行梯度洗脱,以电喷雾离子源负离子多反应监测(MRM)模式进行质谱监测。结果表明:20种化合物可在10 min内完成色谱分离分析,检出限和定量下限分别为0.02~1.59 ng/mL和0.07~5.30 ng/mL,16种黄酮及4种萜类内酯在各自的线性范围内线性关系良好,在低、中、高3个加标水平下的回收率为85.9%~109%。该方法前处理简单、快速高效、准确性高,为舒血宁注射剂的质量控制提供了参考。     

Moringa oleifera hydroethanolic extracts effectively alleviate acetaminophen-induced hepatotoxicity in experimental rats through their antioxidant nature

The aim of the study was to investigate the in vitro antioxidant properties Moringa oleifera Lam. (MO) extracts and its curative role in acetaminophen (APAP)-induced toxic liver injury in rats caused by oxidative damage. The total phenolic content and antioxidant properties of hydroethanolic extracts of different MO edible parts were investigated by employing an established in vitro biological assay. In the antihepatotoxic study, either flowers or leaves extract (200 mg/kg or 400 mg/kg, i.p) were administered an hour after APAP administration, respectively. N-Acetylcysteine was used as the positive control against APAP-induced hepatotoxicity. The levels of liver markers such as alanine aminotransferase (ALT) and the levels of oxidative damage markers including malondialdehyde (MDA), 4-hydroxynonenal (4-HNE) protein adduct, reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) were analysed and compared between experimental groups. Among MO edible parts the flower extracts contain the highest total phenolic content and antioxidant capacity, followed by leaves extract. The oxidative marker MDA, as well as 4-HNE protein adduct levels were elevated and GSH, SOD and CAT were significantly decreased in groups treated with hepatotoxin. The biochemical liver tissue oxidative markers measured in the rats treated with MO flowers and leaves hydroethanolic extracts showed a significant (p < 0.05) reduction in the severity of the liver damage. The results of this study strongly indicate the therapeutic properties of MO hydroethanolic extracts against acute liver injury and thereby scientifically support its traditional use.     

Enhancement by Glycyrrhizae Radix of hepatic metabolism of hypaconitine,a major bioactive and toxic component of Aconiti Laterlis Radix,evaluated by HPLC‐TQ‐MS/MS analysis

Glycyrrhizae Radix (GR) is often prescribed together with Aconiti Laterlis Radix (ALR) (a so‐called compatible drug pair) in traditional Chinese medicinal practice to reduce toxicity of ALR. However, the mechanisms involved remain to be addressed. In this study, the metabolic interactions between GR–ALR drug pair were investigated for the first time. First, an HPLC‐TQ‐MS/MS method was developed to analyze hypaconitine, a major bioactive and toxic component of ALR, in rat liver S9. Then the in vitro metabolic rates of hypaconitine by different rat liver S9 were compared using the established method. The experiments were designed in four groups: pure hypaconitine (group I) and ALR extract (group II) incubated with liver S9 of normal rats, and pure hypaconitine (group III) and ALR extract (group IV) incubated with liver S9 of GR‐pretreated rats. When incubated for more than 4 h, the metabolic rates of hypaconitine in group III were significantly higher than those in group I, and when incubated for more than 2 h, the metabolic rates of hypaconitine in group IV were significantly higher than those in group II, suggesting that GR can enhance metabolic rate of hypaconitine, the mechanism of which might be related to hepatic metabolizing enzyme induction by GR. Copyright © 2012 John Wiley & Sons, Ltd.