糖尿病合并非酒精性脂肪性肝病患者尿代谢组学研究

目的：运用气相色谱-质谱联用技术（GC/MS）技术观察糖尿病合并非酒精性脂肪性肝病（NAFLD）患者尿液中小分子代谢物,发现用于糖尿病合并NAFLD诊断的潜在生物标志物,为糖尿病合并NAFLD的发生机制和早期诊断提供代谢水平上的依据。方法：采用GC/MS检测20例正常人（正常组）、20例单纯糖尿病患者（单纯组）以及30例糖尿病合并NAFLD （合并组）的尿液样本中小分子代谢物。采用主成分分析法（PCA）和偏最小二乘法判别分析（PLS-DA）分析正常组、单纯组以及合并组的代谢谱变化。结果：正常组和单纯组的代谢谱完全分离,正常组和合并组的代谢谱分离良好。利用Wiley和NIST等数据库筛选和鉴定了19个与糖尿病合并NAFLD密切相关的潜在生物标志物。经过代谢通路分析,糖尿病合并NAFLD患者体内存在苯丙氨酸、酪氨酸和色氨酸的生物合成、苯丙氨酸代谢、组氨酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、氨酰tRNA生物合成、色氨酸代谢、酪氨酸代谢等代谢异常。结论：糖尿病合并NAFLD的发生与苯丙氨酸、酪氨酸和色氨酸的生物合成、苯丙氨酸代谢、组氨酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、氨酰tRNA生物合成、色氨酸代谢、酪氨酸代谢等代谢异常有关,尿液中鉴定的代谢物有望成为糖尿病合并NAFLD诊断的潜在生物标志物。     

从水通道蛋白的角度探讨蚕沙的“化湿”作用机制

目的：从水通道蛋白角度研究蚕沙的"化湿"作用。方法：SD大鼠，随机分为5组，即正常组、模型组、蚕沙低、中、高剂量组。采用"外湿侵体+正气耗损+过食肥甘"的方法，建立"湿阻中焦"证模型，Western blot法检测肾AQP1、肺AQP1、结肠AQP3、皮肤AQP3、颌下腺AQP5蛋白的表达，qPCR法检测AQP mRNA的表达。结果：与正常组比较，模型组大鼠肾中AQP1蛋白和mRNA表达明显升高（P<0.05），且肺AQP1、结肠AQP3、皮肤AQP3、颌下腺AQP5蛋白和mRNA表达均明显降低（P<0.05）；与模型组比较，蚕沙高剂量组肾AQP1蛋白和mRNA表达显著降低（P<0.05），肺AQP1、结肠AQP3、皮肤AQP3、颌下腺AQP5蛋白和mRNA表达均显著升高（P<0.05）。结论：蚕沙可能通过调节肾、肺、结肠、皮肤、颌下腺组织中AQP的表达，发挥"化湿"的作用。     

慢性阻塞性肺病稳定期模型大鼠支气管肺泡灌洗液的代谢组学研究

目的：基于代谢组学方法研究慢性阻塞性肺病（chronic obstructive pulmonary disease，COPD）稳定期大鼠支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）的潜在生物标志物。方法：采用烟熏联合细菌反复感染的方法制备COPD稳定期模型大鼠。采用超高效液相色谱-串联质谱技术采集大鼠的BALF数据，利用多元统计分析软件筛选差异代谢物，再结合数据库鉴定潜在生物标志物，并进行代谢通路分析。结果：多元统计分析结果显示，正常组与模型组BALF代谢物发生显著变化。通过比对数据库，共鉴定出30个差异性代谢物，主要涉及到体内的氨酰基-tRNA的生物合成，甘氨酸、丝氨酸和苏氨酸的代谢，鞘脂代谢，苯丙氨酸代谢和花生四烯酸代谢等8条代谢通路。结论：基于代谢组学研究COPD稳定期的BALF代谢物变化，寻找COPD稳定期相关代谢标志物，为治疗COPD提供理论参考。     

糖尿病周围神经病变患者尿代谢组学研究

目的：采用气相色谱-质谱联用技术（GC/MS）技术,分析糖尿病周围神经病变患者尿液中小分子代谢物,为糖尿病周围神经病变的发生机制和早期诊断提供代谢水平上的依据。方法：采用气相色谱-质谱联用技术（GC/MS）检测40例正常人（正常组）、20例单纯糖尿病患者（单纯组）以及40例糖尿病周围神经病变患者（合并组）的尿液样本中小分子代谢物。采用主成分分析法（PCA）和偏最小二乘法判别分析方法（PLS-DA）观察正常组、单纯组以及合并组的代谢谱的变化。结果：正常组和单纯组的代谢谱区分良好,正常组和合并组的代谢谱也能良好的区分。利用Wiley和NIST等数据库筛选和及本实验室建立的标准品代谢物谱库,鉴定了17个与糖尿病周围神经病变密切相关的潜在生物标志物。经过代谢通路分析,糖尿病周围神经病变患者体内存在牛磺酸和亚牛磺酸代谢、甘氨酸、丝氨酸和苏氨酸代谢等代谢异常。结论：糖尿病周围神经病变发生与牛磺酸和亚牛磺酸代谢、甘氨酸,丝氨酸和苏氨酸代谢等代谢异常有关,尿液中鉴定的代谢物有望成为糖尿病周围神经病变诊断的潜在生物标志物。     

黄芪甲苷对阿德福韦酯致肾损伤的保护作用和机制研究

目的：本实验旨在研究黄芪甲苷（astragaloside IV,As）对阿德福韦酯（adefovir dipivoxil,Adv）致肾损伤的保护作用,探究其可能的作用机制。方法：将50只SD雄性大鼠按数字表法随机分为正常组（normal）,Adv肾损伤组（30 mg·kg^-1·d^-1）,As低、中、高剂量干预组（10,20,30 mg·kg^-1·d^-1）。通过测定血浆生化指标比较各组肾功能差异;Masson染色评价各组肾小管上皮细胞病理学改变;利用UHPLC-MS代谢组学方法检测各组大鼠血浆内源性代谢物的变化,筛选出差异代谢物。结果：与正常组相比,Adv组肌酐值显著升高,肾损伤严重。而As可显著降低肌酐值和改善肾小管上皮细胞病理学形态,有效干预Adv致肾损伤。Adv组大鼠血浆共发现28个差异代谢物,As可显著逆转16个代谢物的变化。主要涉及与能量代谢相关的苯丙氨酸、酪氨酸和色氨酸代谢通路、泛醌和其他萜烯-醌代谢通路,与氧化应激相关的谷胱甘肽代谢通路。结论：本研究提示As保护Adv致肾损伤的机制与改善细胞供能,减少氧化应激对细胞的损害密切相关。本研究为阿德福韦酯治疗慢性乙型肝炎（CHB）时利用黄芪作为减毒增效的治疗策略提供了基础依据。     

基于气相色谱-质谱联用技术的肥胖儿童血清代谢组学研究

目的：筛选与儿童肥胖相关的血液中差异代谢物,探讨肥胖儿童代谢紊乱的发生机制。方法：采用气相色谱-质谱联用（GC/MS）和模式识别技术分析肥胖儿童血清中代谢物的变化,GC/MS检测25例肥胖儿童（肥胖组）和25例健康儿童（正常组）血清中代谢物,采用偏最小二乘判别分析（partial least squares discrimination analysis,PLS-DA）识别两组样本的血清代谢谱及代谢物。结果：肥胖组和正常组的代谢谱分离良好,肥胖儿童和正常儿童之间的血清代谢成分存在33个差异代谢物,绘制33个差异代谢物ROC曲线,计算曲线下面积,肌酐、马来酸、莽草酸等16个代谢物面积大于0.5,可能是诊断肥胖儿童潜在的生物标志物。结论：肥胖儿童患者血清代谢物的变化机制和甘氨酸,丝氨酸和苏氨酸的代谢、丙氨酸、天冬氨酸和谷氨酸代谢、乙醛酸和二羧酸酯代谢密切相关。     

基于血浆代谢组学评价不同神经保护剂联用方案对缺血性脑卒中生物标志物的影响

目的：通过代谢组学挖掘神经保护剂联用治疗缺血性脑卒中患者血浆的回调性生物标志物,从代谢路径角度评价神经保护剂联用的合理性。方法：通过高效液相色谱与飞行时间质谱（Q-TOF）联用的非靶向代谢组学平台,获取缺血性脑卒中患者的血浆代谢组学数据,用多元变量统计分析进行数据处理,筛选出药物干预后回调的生物标记物及其代谢路径。结果：代谢组学研究表明缺血性脑卒中患者体内主要呈氨基酸、脂类及能量代谢紊乱,给予不同神经保护剂联用方案后整体代谢轮廓回调,经数据库鉴定和匹配后得到两药联用-丁苯酞+依达拉奉组、两药联用-丁苯酞+单唾液酸组和三药联用-丁苯酞+依达拉奉+单唾液酸组显著变化的共性回调生物标记物及特征性回调生物标记物（溶血磷脂酰胆碱18∶0、色氨酸和苹果酸等）。结论：神经保护剂联合应用可使缺血性脑卒中患者的代谢紊乱恢复。调节能量代谢（三羧酸循环）、脂类代谢（甘油磷脂代谢、脂肪酸代谢）和氨基酸代谢（芳香族氨基酸代谢、鸟氨酸代谢）通路等可能是其治疗缺血性脑卒中的神经保护机制。     

终末期肾衰竭维持血液透析患者的血清代谢组学研究

目的：采用气相色谱-质谱联用（GC/MS）为基础的代谢组学技术分析终末期肾病（end stage renal disease,ESRD）维持血液透析患者血清中代谢物的变化,筛选出可用于ESRD早期临床诊断的潜在生物标志物,探讨ESRD血液透析患者代谢紊乱的发生机制。方法：采用GC/MS检测63例ESRD患者（病例组）和相匹配的20例健康人（对照组）血清中代谢物,采用偏最小二乘判别分析法（partial least squares discrimination analysis,PLS-DA）分析两组样本血清代谢谱,寻找两组之间的差异代谢物。结果：对照组和病例组的代谢谱区分良好,利用数据库NIST、Wiley Registry代谢组数据库和标准品数据库鉴定出34个差异代谢物,绘制34个差异代谢物ROC曲线,计算曲线下面积,羟胺、尿素、富马酸等28个代谢物面积大于0.5。结论：ESRD患者存在能量代谢、氨基酸代谢、脂质代谢、尿素循环、嘌呤代谢等代谢异常,血清中鉴定的羟胺、尿素、富马酸等28个代谢物有望成为诊断ESRD的潜在生物标志物。     

20(R)-25-OCH_3-PPD在大鼠体内代谢产物的鉴定

目的鉴定抗肿瘤药物20(R)-25-OCH3-PPD在大鼠体内的代谢产物。方法大鼠单次灌胃给予药物20(R)-25-OCH3-PPD(40 mg·kg-1),收集粪便、尿液及血浆。采用LC-MS/MS法,以多反应监测(MRM)和二级全扫描质谱(Full Scan MS2)方式,对大鼠粪便、尿液及血浆中代谢产物进行分析和鉴定。结果在粪中检测到原形药物及8个代谢物,在血浆中检测到原形药物和1个去甲基代谢物,在尿液中没有检测到相关代谢物。结论 20(R)-25-OCH3-PPD在大鼠体内代谢广泛。     

基于序贯代谢模型研究地榆鞣质在大鼠体内的代谢作用

目的：以序贯代谢模型为基础,研究地榆鞣质成分在大鼠体内代谢动态情况。方法：采用超高液相色谱-线性离子阱-串联静电轨道阱高分辨质谱（UHPLC-LTQ-Orbitrap HRMS）技术,以此同时检测多成分变化情况。体内代谢采用在体封闭肠环法研究地榆鞣质口服后多成分经肝肠代谢变化过程。结果：口服地榆鞣质通过肝肠的代谢,经过水解、甲基化及葡萄糖醛酸化后共鉴定出12种代谢产物。结论：采用序贯代谢模型的方法,研究地榆鞣质口服经不同代谢部位的情况进行了定性描述,可为该药物的质量控制、机制阐述及再次开发提供参考依据。     

The metabolomic profiling of serum in rats exposed to arsenic using UPLC/Q-TOF MS

Chronic arsenicosis induced by excessive arsenic intake can cause damages to multi-organ systems, skin cancer and various internal cancers. However, the key metabolic changes and biomarkers which can reflect these changes remain unclear resulting in a lack of effective prevention and treatments. The aim of this study is to determine the impact of chronic arsenic exposure on the metabolism of organism, and find the metabolites changes by using metabolomic techniques. Thirty male Wistar rats were randomly divided into three groups. The arsenite was administered in water, and the doses were 0, 10, and 50 mg/L, respectively. The exposure lasted for 6 months. The endogenous metabolite profile of serum was investigated by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Partial least squares discriminant analysis (PLS-DA) enabled clusters to be visualized. Nine serum principal metabolites contributing to the clusters were identified, which were CPA (18:2(9Z,12Z)/0:0), LysoPC (14:0), LysoPC (18:4 (6Z,9Z,12Z,15Z)), LysoPC (P-18:0), l-palmitoylcarnitine, LysoPC (20:2(11Z,14Z)) in positive ESI mode and deoxygcholylglycine, LysoPE (0:0/20:2(11Z,14Z)), 15(S)-hydroxyeicosatrienoic acid in negative ESI. These changes of metabolites in rats suggested the changed metabolism in rats exposed to arsenic. These findings may further aid diagnose and serve as targets for therapeutic intervention of arsenicosis.     

Metabolomics combined with serum pharmacochemistry discovering effective compounds of Fangji Huangqi Tang against nephrotic syndrome

OBJECTIVE An integrative strategy was established to discover the effective compounds and their therapeutic targets using Fangji Huangqi Tang(FHT) aiming at inhibiting nephrotic syndrome(NS) as a case study. METHODS The adriamycin-induced nephropathy rat model was evaluated by histopathology analysis and urine protein. The serum biomarkers(pathological marker)related to NS model were characterized by metabolomics, and the metabolites which could be regulated to normal levels after administration with FHT were defined as FHT-regulated biomarkers(effective marker). Moreover, the potential effective compounds were identified by comparison of drug serum between control and model rats. Furthermore, they were further screened based on the correlation analysis between effective marker with the potential effective compounds. At the same time, the potential target of effective ingredients was found by network pharmacology technology. RESULTS The results of serum metabonomics showed that 15 metabolites,including 3-Hydroxybutyric acid, L-phenylalanine and linolenic acid, were associated with renal damage. Among them, 6 effective markers were uric acid, 2-methylbutyrylcarnitine and 10-HDA. Metabolic pathway analysis showed that, phenylalanine, tyrosine and tryptophan biosynthesis, linoleic acid metabolism, phenylalanine metabolism, sphingolipid metabolism were the key pathway associated with NS. The correlation analysis showed that nine constituents such as fan Ghinoline,atractylenolide Ⅲ, cycloastragenol, glycyrrhetinic acid were recognized as effective compounds, whose potential protein targets participated in MAPK signaling pathway,Gn RH signaling pathway and aldoaterone-regulated sodium reabsorption. CONCLUSION This study provides a methodological reference for the study of the efficacy material base of other traditional Chinese medicineand also provides an important basis for the target of FHT against NS.     

UPLC-Q/TOF-MS based metabonomics revealed protective effect of Terminalia chebula extract on ischemic stroke rats

Terminalia chebula(TC), a kind of Combretaceae, is a widely used herb in India and East Asia to treat cerebrovascular diseases. However, the potential mechanism of the neuroprotective effects of TC at the metabonomics level is still not unclear. The present study focused on the effects of TC on metabonomics in stroke model. In our study,rats were divided randomly into Sham, Model, and TC groups. The TC group were intragastricly administered with TC for 7 d after middle cerebral artery occlusion(MCAO) operation. The sham and the model groups received vehicle for the same length of time. Subsequently, the neuroprotective effects of TC were examined by neurological defects evaluation, infarct volume assessment, and identification of biochemical indicators for antioxidant and anti-inflammatory activities. Further, metabonomics technology was employed to evaluate the endogenous metabolites profiling systematically.Consist to results of biochemical and histopathological assays, pattern recognition analysis showed a clear separation of the Model and the Sham group, indicating a recovery impact of TC on the MCAO rats. Moreover, 12 potential biomarkers were identified in MCAO Model group, involved in energy(lactic acid, succinic acid, and fumarate), amino acids(leucine,alanine, and phenylalanine) and glycerophospholipid [PC(16∶0/20∶4), PC(20:4/20:4), Lyso PC(18:0) and Lyso PC(16:0)]metabolism, and other types of metabolism(arachidonic acid and palmitoylcarnitine). Notably, we found that metabolite levels of TC group were partially reversed to normal. In conclusion, TC could ameliorate MCAO rats by intervening with energy metabolism(glycolysis and TCA cycle), amino acid metabolism, glycerophospholipid metabolism and other types of metabolism.     

基于血浆代谢组学的新生化颗粒对急性血瘀大鼠作用机制研究

目的 研究新生化颗粒对急性血瘀大鼠血浆中内源性代谢物的影响,探讨新生化颗粒治疗急性血瘀证的可能机制。方法 将50只SD大鼠随机分为对照组、模型组、复方丹参滴丸（阳性药,0.10 g·kg^-1）组和新生化颗粒低、高剂量（4.86、9.72 g·kg^-1）组,给药体积10 mL·kg^-1,对照组和模型组大鼠ig给予等体积0.5%羧甲基纤维素钠（CMC-Na）,每天早晚各ig给药1次,共7次。第5次给药后,除对照组外,采用sc盐酸肾上腺素和冰水浴制备大鼠急性血瘀模型。通过测定全血黏度（WBV）、血浆黏度（PV）、活化部分凝血活酶时间（APTT）、凝血酶原时间（PT）、纤维蛋白原（FIB）和凝血酶时间（TT）,观察不同剂量的新生化颗粒对急性血瘀大鼠血液流变学和凝血功能的影响;采用超高效液相色谱-四极杆飞行时间质谱联用（UHPLCQTOF-MS）法检测各组大鼠血浆中的内源性代谢物,通过多变量统计分析筛选潜在生物标志物,结合质谱信息、数据库检索和标准品比对鉴定潜在生物标志物,并将鉴定到的生物标志物导入MetaboAnalyst 5.0数据库推测其可能的代谢通路。结果 与模型组比较,新生化颗粒显著降低急性血瘀大鼠WBV、PV和FIB,显著延长APTT、PT和TT（P＜0.05、0.01）。代谢组学结果显示,从急性血瘀大鼠血浆中共鉴定出21个差异代谢物（准确鉴定7个）,与对照组比较,模型组大鼠血浆中乳酸、肉碱和肌酐等10个内源性代谢物显著上调,苹果酸、琥珀酸和色胺等11个内源性代谢物显著下调（P＜0.05、0.01）;除了硫酸吲哚酚和脱氧胞苷外,新生化颗粒对其他19个生物标志物均有显著回调作用（P＜0.05、0.01）。这些标志物主要涉及苯丙氨酸、酪氨酸和色氨酸生物合成、苯丙氨酸代谢、亚油酸代谢、三羧酸循环和色氨酸代谢。结论 新生化颗粒对急性血瘀大鼠体内紊乱代谢物有较好的回调作用,其作用机制主要与调节苯丙氨酸、酪氨酸和色氨酸生物合成,苯丙氨酸代谢,亚油酸代谢,三羧酸循环和色氨酸代谢通路有关。     

The effect of prolonged sodium phenobarbitone treatment on hepatic xenobiotic metabolism and the urinary excretion of metabolites of the d-glucuronic acid pathway in the rat

Male Sprague-Dawley rats were fed a 0.1 % (w/w) sodium phenobarbitone (PB) diet for periods of 4, 12 and 24 weeks. Phenobarbitone treatment resulted in a marked increase in liver size which was accompanied by the induction of several parameters of hepatic xenobiotic metabolism including mixed function oxidase enzyme activities, UDP-glucuronyltransferase, cytochrome P450 and the microsomal protein content. In addition, treatment with the barbiturate led to the stimulation of the urinary excretion of d-glucaric acid, l-gulonic acid, free d-glucuronic acid, l-ascorbic acid and xylitol. The levels of stimulation of both the hepatic and urinary parameters remained relatively constant throughout the period of treatment. On cessation of PB treatment the parameters of hepatic xenobiotic metabolism reverted to control levels at a faster rate than the excretion of the d-glucuronic acid metabolites. The results demonstrate a correlation between the urinary excretion of metabolites of the d-glucuronic acid pathway and hepatic xenobiotic metabolizing enzyme activities during the prolonged administration of PB.     

基于GC-TOF/MS技术分析肾性高血压大鼠血清代谢组学变化

目的 基于气相色谱-质谱（GC-TOF/MS）技术分析两肾一夹（2K1C）肾性高血压大鼠血清代谢物的变化 情况。方法 20只健康雄性SD大鼠按照随机数字表法分为假手术（Sham）组和2K1C组,每组10只。比较建模前及 建模4周后2组大鼠体质量及尾动脉收缩压变化。采用GC-TOF/MS检测血清代谢产物的表达水平,正交偏最小二乘 法-判别分析（OPLS-DA）筛选出差异表达代谢物,并对筛选出的差异表达代谢物进行聚类分析及KEGG通路分析。 结果 4周后2K1C组共7只大鼠建模成功。与Sham组相比,2K1C组共筛选到14种差异表达代谢物,其中,花生四 烯酸（arachidonic acid）、马尿酸（hippuric acid）、七烷酸（heptadecanoic acid）、吲哚乳酸酯（indolelactate）、木糖（xylose）、 顺-巨头鲸鱼酸（cis-gondoic acid）、富马酸（fumaric acid）、胞苷-磷酸（cytidine-monophosphate）、乳酰胺（lactamide）、2- 羟基-3-异丙基丁二酸（2-hydroxy-3-isopropylbutanedioic acid）、双缩脲（biuret）这 11 种代谢物表达上调,酪氨酸 （tyrosine）、胆酸（cholic acid）、豆甾醇（stigmasterol）表达下调。以上差异表达代谢物与能量代谢、氨基酸代谢、脂质代 谢、初级胆汁酸生物合成等生物过程密切相关。结论 2K1C肾性高血压大鼠血清代谢物发生变化,并涉及能量代谢 等多种通路,进而影响肾性高血压的病理生理过程。     

Metabolism of doxylamine succinate in Fischer 344 rats. Part II: Nonconjugated urinary and fecal metabolites

Elimination and metabolic profiles of doxylamine and its nonconjugated metabolites were determined after the oral administration of [14C]-doxylamine succinate (13.3 mg/kg and 133 mg/kg doses) to male and female Fischer 344 rats. Total urine and fecal recovery of the administered dose was greater than 90% regardless of sex or dose. The cumulative urinary and fecal elimination of these nonconjugated doxylamine metabolites at the 13.3 mg dose was 44.4 +/- 4.4% and 36.0 +/- 5.8% of the total recovered dose for male and female rats, respectively. The cumulative urinary and fecal elimination of the doxylamine nonconjugated metabolites at the 133 mg/kg dose was 38.7 +/- 2.7% and 41.4 +/- 1.0% of the total recovered dose for male and female rats, respectively. In order to determine the contribution of mammalian and bacterial enzymes in the overall metabolism and excretion patterns for doxylamine, two in vitro techniques were investigated. Incubation of [14C]-doxylamine succinate with human and rat intestinal microflora indicated that anaerobic bacteria were not capable of effecting the degradation of [14C]-doxylamine succinate. However, the incubation of [14C]-doxylamine succinate with isolated rat hepatocytes generated several metabolites similar to those observed in vivo. The nonconjugated doxylamine metabolites isolated and identified include: doxylamine N-oxide, desmethyldoxylamine, didesmethyldoxylamine and ring-hydroxylated products of doxylamine and desmethyldoxylamine. The studies demonstrate the role of hepatic metabolism in the elimination of doxylamine succinate in the rat.     

基于尿液代谢组学研究碘克沙醇损伤肾小管功能的作用机制

目的 研究使用碘克沙醇行冠脉造影后肾小管功能出现损伤患者尿液中内源性代谢物的代谢特征，分析碘克沙醇导致肾小管损伤的潜在机制。方法 选择使用碘克沙醇冠脉造影后24 h内肾小管功能正常和肾小管功能出现损伤的患者各50例，分为正常组(n=50)和损伤组(n=50)，收集其造影24 h内的晨尿尿液样本。采用超高效液相色谱串联质谱以及正交偏最小二乘法(orthogonal partial least squares，OPLS-DA)对尿液样品进行代谢分析；利用Metlin、HMDB、KEGG、MetaboAnalyst数据库考察碘克沙醇损伤肾小管功能的代谢特征。结果 鉴定出与碘克沙醇损伤肾小管功能相关的差异代谢物25种[变量权重值(variable important in projection，VIP)>1]，其中有3种代谢差异代谢物的含量显著下降(VIP>1，P<0.05)，分别为L-苯丙氨酸、L-酪氨酸、L-色氨酸。25种差异代谢物涉及的代谢通路有9条，主要为氨基酸代谢，其中Impact>0.10，P<0.05的目标代谢通路有2条，分别为苯丙氨酸、酪氨酸和色氨酸的生物合成通路及苯丙氨酸代谢通路。结论 使用碘克沙醇行冠脉造影后肾小管功能出现损伤患者的尿液代谢主要表现为氨基酸代谢紊乱，尿液中L-酪氨酸、L-苯丙氨酸、L-色氨酸含量显著下降可提示肾小管功能出现损伤。碘克沙醇相关早期肾损伤的发病机制主要是通过影响苯丙氨酸、酪氨酸和色氨酸生物合成通路，苯丙氨酸代谢通路来损伤肾小管，进而影响其功能。     

Evaluation of Cadmium-Induced Nephrotoxicity Using Urinary Metabolomic Profiles in Sprague-Dawley Male Rats

The aim of this study was to investigate urinary metabolomic profiles associated with cadmium (Cd)-induced nephrotoxicity and their potential mechanisms. Metabolomic profiles were measured by high-resolution ¹H-nuclear magnetic resonance (NMR) spectroscopy in the urine of rats after oral exposure to CdCl₂ (1, 5, or 25 mg/kg) for 6 wk. The spectral data were further analyzed by a multivariate analysis to identify specific urinary metabolites. Urinary excretion levels of protein biomarkers were also measured and CdCl₂ accumulated dose-dependently in the kidney. High-dose (25 mg/kg) CdCl₂ exposure significantly increased serum blood urea nitrogen (BUN), but serum creatinine (sCr) levels were unchanged. High-dose CdCl₂ (25 mg/kg) exposure also significantly elevated protein-based urinary biomarkers including osteopontin, monocyte chemoattractant protein-1 (MCP-1), kidney injury molecules-1 (Kim-1), and selenium-binding protein 1 (SBP1) in rat urine. Under these conditions, six urinary metabolites (citrate, serine, 3-hydroxyisovalerate, 4-hydroxyphenyllactate, dimethylamine, and betaine) were involved in mitochondrial energy metabolism. In addition, a few number of amino acids such as glycine, glutamate, tyrosine, proline, or phenylalanine and carbohydrate (glucose) were altered in urine after CdCl₂ exposure. In particular, the metabolites involved in the glutathione biosynthesis pathway, including cysteine, serine, methionine, and glutamate, were markedly decreased compared to the control. Thus, these metabolites are potential biomarkers for detection of Cd-induced nephrotoxicity. Our results further indicate that redox metabolomics pathways may be associated with Cd-mediated chronic kidney injury. These findings provide a biochemical pathway for better understanding of cellular mechanism underlying Cd-induced renal injury in humans.     

Tissue distribution and urinary excretion of inorganic arsenic and its methylated metabolites in C57BL6 mice following subchronic exposure to arsenate in drinking water

The relationship of exposure and tissue concentration of parent chemical and metabolites over prolonged exposure is a critical issue for chronic toxicities mediated by metabolite(s) rather than parent chemical alone. This is an issue for AsV because its trivalent metabolites have unique toxicities and relatively greater potency compared to their pentavalent counterparts for many endpoints. In this study, dose-dependency in tissue distribution and urinary excretion for inorganic arsenic and its methylated metabolites was assessed in female C57Bl/6 mice exposed to 0, 0.5, 2, 10 or 50 ppm arsenic (as arsenate, AsV) in their drinking water for 12 weeks. No adverse effects were observed and body weight gain did not differ significantly among groups. Urinary excretion of arsenite monomethylarsonous acid (MMA(III)), dimethylarsinous acid (DMA(III)), dimethylarsinic acid (DMAV), and trimethylarsine oxide (TMAO) increased linearly with dose, whereas AsV and monomethylarsonic acid (MMAV) excretion was non-linear with respect to dose. Total tissue arsenic accumulation was greatest in kidney > lung > urinary bladder > skin > blood > liver. Monomethyl arsenic (MMA, i.e. MMA(III)+MMAV) was the predominant metabolite in kidney, whereas dimethylarsenic (DMA, i.e., DMA(III)+DMAV) was the predominant metabolite in lung. Urinary bladder tissue had roughly equivalent levels of inorganic arsenic and dimethylarsenic, as did skin. These data indicate that pharmacokinetic models for arsenic metabolism and disposition need to include mechanisms for organ-specific accumulation of some arsenicals and that urinary metabolite profiles are not necessarily reflective of target tissue dosimetry.