硝酸镥急性毒性的体液核磁共振氢谱研究

采用现代核磁共振技术,通过分析灌胃给药0．01、0．05、0．2、2、10和100ms／kg剂量Lu(NO3)3 24h内大鼠尿液及24h后大鼠血清的核磁共振氢谱(^1HNMR),由体液中内源性代谢物浓度的变化研究了稀土化合物在动物体内急性毒性。较高剂量组体液中的氨基酸、尿囊素、柠檬酸、氮氧三甲胺和肌酸酐等重要内源性代谢物的核磁共振谱峰强度发生了明显的变化,说明动物体内的代谢物出现异常：高剂量的稀土的引入可能使动物肾脏和肝脏均受到损害,且受损程度随稀土剂量的增高而渐趋严重。     

《稀土生物无机化学》(第2版)征订启事

《稀土生物无机化学》(第 2版 )是由倪嘉缵院士 ,王夔院士 ,杨频教授等国内长期从事稀土生物无机化学的同志撰写。该书在第 1版的基础上 ,增加了许多新的内容 ,其中绝大部分是国家自然科学基金委员会化学科学部及生命科学部资助的重大研究项目的最新研究成果。许多章节是从基因水平 ,分子水平 ,细胞水平 ,直至动物的整体试验对稀土进入动物和植物体内的物种分布 ,垮红细胞膜的输运 ,稀土对核酸的作用 ,对体内的一些主要蛋白的相互作用 ,如血红蛋白 ,运铁蛋白 ,钙调蛋白等 ,及稀土进入动物体后对内分泌体系的影响等作了系统的论述。故可供从…     

我国稀土生物无机化学研究进展

我国稀土生物无机化学研究的特点是紧密结合稀土在农用及药物中应用所提出急需回答的问题, 经过长期的工作, 已在稀土的跨细胞膜运转, 稀土进入动物体内后的物种分布, 稀土与细胞膜的作用, 稀土对血红蛋白结构与载氧功能的作用, 稀土对骨细胞及骨结构的影响以及稀土在大鼠肝脏中的积累等方面取得了满意的结果. 从而可以为今后开展稀土元素在农用及药用中危险性评价的基础数据, 同时亦将促进稀土细胞化学的发展.     

稀土对大鼠代谢产物影响的核磁共振研究

以高场核磁共振技术为研究手段,通过分析腹腔注射０．２、２、１０、２０ｍｇ／ｋｇ体重剂量的Ｌａ（ＮＯ３）３后大鼠尿液中代谢物浓度、物种的变化,研究了稀土化合物在动物体内的作用情况,结果表明,稀土的引入使动物肾脏和肝脏都受到一定程度的损伤,并在代谢物中挑选出了合适的ＮＭＲ ｍａｒｋｅｒｓ,其变化可以反映稀土离子作用后大鼠的异常代谢。     

量热法测定氯化钐与甘氨酸配合物的标准生成焓

稀土在生命科学领域的研究日益受到人们关注,稀土化合物所具有的抑菌、抑癌、消炎等作用及其作用机理的探讨已有报道［１～３］。最近几年稀土在生物领域中的研究又有了新的突破和进展,从稀土与氨基酸、蛋白质、膜脂及膜蛋白的作用到其对ＤＮＡ。ＲＮＡ的影响［１,４］,从稀土的分子水平、细胞及亚细胞水平到动物整体实验的系统研究［５,６］等,分别从不同的层次、不同的水平研究了稀土的生物效应。但至目前为止,稀土的生物效应机理及其对人体的影响尚未得到令人满意的解释。由于氨基酸是构成人体蛋白质的基本单位,故研究稀土与氨基…     

合成化合物动物体内抗肿瘤实验方法研究进展

进行了动物体内抗肿瘤活性研究的多为天然药物,但其来源有限;虽然用于抗肿瘤研究的合成化合物种类较多,但对于其在动物体内抗肿瘤研究又相对偏少,所以,合成化合物抗肿瘤体内研究已经成为研究热点之一。本文对合成化合物抗肿瘤作用机制、体内肿瘤模型建立方法和体内抑瘤实验研究等方面的研究进展进行了评述,并通过大量实验数据对合成化合物毒性评价方法、抑瘤基本观察指标测定、氧化损伤相关酶活力的测定方法、组织病理学评价方法等体内抑瘤实验的评价标准和方法进行了详细总结,提出了合成化合物在体内动物抗肿瘤实验研究方法中亟待解决的问题,为新型抗肿瘤合成化合物体内药学活性研究提供有益的参考。     

硝酸铈急性生物效应的肝肾组织魔角旋转NMR代谢组学研究

采用现代核磁共振技术中魔角旋转方法,通过分析腹腔注射给药2,10和50 mg.kg-1体重剂量硝酸铈48 h后大鼠肝和肾组织MAS1H NMR谱,由组织中内源性代谢物浓度的变化研究了硝酸铈在动物体内的急性毒性,并利用模式识别技术对硝酸铈急性生物效应的NMR marker进行了研究。较高剂量组肝组织中的甘油三酯、乳酸、肝糖原和肾组织中的甘油三酯、甘油磷酸胆碱、甜菜碱等重要内源性代谢物的核磁共振谱峰强度发生了明显的变化,意味着动物体内的代谢出现异常:高剂量硝酸铈的引入可使动物肾脏和肝脏均受到损害,且受损程度随稀土剂量的增高而渐趋严重。     

纳米材料生物效应及其毒理学研究进展

纳米科学与信息科学和生命科学并列, 已经成为21世纪的三大支柱科学领域. 由于纳米材料独特的物理化学性质, 纳米尺度及纳米结构的材料乃至器件, 已逐渐走出实验室, 进入人们的生活. 这些具有独特物理化学性质的纳米材料, 对人体健康以及环境将带来的潜在影响, 目前已经引起科学界, 乃至政府部门的广泛关注. 文中分析综述了几种纳米材料(纳米TiO₂、单壁碳纳米管、多壁碳纳米管及超细铁粉)目前已取得的部分生物效应及毒理学的研究结果, 包括纳米材料在生物体内的分布、作用的靶器官、纳米材料引起的细胞毒性、细胞凋亡等. 文中还评价了纳米颗粒的生物毒性. 纳米颗粒的尺寸越小, 显示出生物毒性的倾向越大; 尽管碳纳米管是由石墨层卷成的圆筒, 但是根据石墨的安全剂量来外推碳纳米管的安全剂量是不可行的, 碳纳米管的生物毒性远大于石墨粉; 表观分子量高达60万的水溶性纳米碳管, 在小鼠体内却显示出小分子的生理行为; 一种正在研究的磁性纳米颗粒在动物体内显示出迅速团聚、堵塞血管等现象. 纳米材料在生物体内呈现出的这些生理现象, 仅利用现有的知识尚无法解释. 最后还介绍了纳米物质生物效应(包括毒理学, 安全性)研究的部分实验方法; 展望了该新领域今后的发展方向和亟待研究的重要问题.     

从稀土对骨代谢的影响看稀土的药用及安全性

骨骼的正常代谢涉及多种金属离子,随着稀土的应用,通过环境和经由食物链等渠道进入人体内的稀土量也有所增加, 并且在骨组织中相对富集且难排出;由于稀土与钙离子的相似性,它们将干预和调控骨的形成与再造。一方面,稀土被设想可以用来作为治疗骨疾病的药物, 另一方面,它对骨代谢的影响产生负面作用。因此, 稀土的药用和安全性日益成为人们关注的问题。本文回顾了稀土离子对骨代谢影响的研究进展,探讨了其药用和安全性问题, 主要从以下三个方面进行综述: (1)稀土离子对成骨细胞细胞和破骨细胞的分化和功能表达的影响; (2)稀土离子对骨髓基质细胞成骨分化和成脂分化的影响; (3)稀土离子对动物骨矿物相的影响。此外,还指出了该研究领域的不足以及今后应该加强的研究方向。     

稀土发挥生物效应的机制及其作用物种探讨

稀土在工业、医药领域、基础研究以及在我国的广泛农用引起了人们对其生物效应机理以及可能毒性的关注.在稀土生物学效应机理及毒性的研究中,无论是在动物水平还是细胞层次,引起生物学效应的稀土物种都是一个关键问题,一直存在争议.本文对以动物、细胞为模型的生物效应研究中的实验条件进行分析,对生理条件下引起稀土生物学效应的可能物种提出"稀土离子池"(rare earth ion pool)模型,并对其引起生物学效应的活性物种以及与细胞膜相互作用的方式进行了探讨,以期为阐明复杂生物学体系中稀土化合物的作用机制提供思路.     

Evaluation of Bioeffects of a Long-term Administering ChangLe on Rat by Proton NMR Method and BIochemical Examination

High resolution ¹H nuclear magnetic resonance (NMR) spectroscopy has been employed to assess long‐term toxicological effects of ChangLe (a kind of rare earth complex applied in agriculture). Male Wistar rats were administrated orally with ChangLe at doses of 0, 0.1, 0.2, 2.0, 10 and 20 mg/kg body weight daily, respectively, for 6 months. Urine was collected at day 30, 60, 90 and serum samples were taken after 6 months. Many low‐molecular weight metabolites were identified by ¹H NMR spectra of rat urine. A decrease in citrate and an increase in ketone bodies, creatinine, DMA, DMG, TMAO, and taurine in the urine of the rats receiving high doses were found by ¹H NMR spectra. These may mean that high‐dosage of ChangLe impairs the specific region of liver and kidney, such as renal tubule and mitochondria. The decrease in citrate and the increase in succinate and α‐ketoglutarate were attributed to a combination of the inhibition of certain citric acid enzymes, renal tubular acidosis and the abnormal fatty acid catabolism. The information of the renal capillary necrosis could be derived from the increase in DMA, DMG and TMAO. The increase in taurine was due to hepatic mitochondria dysfunction. The conclusions were supported by the results of biochemical measurements and enzymatic assay. No obviously toxicological effects on metabolism were found for low‐dosage ChangLe, but further studies on safety of ChangLe applied to agriculture are still required.     

Urine 2DE proteome analysis in healthy condition and kidney disease

Urine is a source of potential markers of disease. In the context of renal disease, urine is particularly important as it may directly reflect kidney injury. Current markers of renal dysfunction lack both optimal specificity and sensitivity, and improved technologies and approaches are needed. There is no clear consensus about the best sample pretreatment procedure for 2DE analysis of the urine proteome. Sample pretreatment conditions spots resolution and detection sensitivity, critically. As a first goal, we exhaustively compared eight different sample cleaning and protein purification methodologies for 2DE analysis of urine from healthy individuals. Oasis® HLB cartridges allowed the detection of the highest number of low molecular weight proteins; while PD10 desalting columns resulted in the highest number of detected spots in the high molecular weight area. Sample pretreatment strategies were also explored in the context of proteinuria, a clinical condition often associated to renal damage. Testing of urine samples from 13 patients with hypertension or kidney disease and different levels of proteinuria identified Oasis® HLB cartridge purification in combination with albumin depletion by ProteoPrep kit as the best option for urine proteome profiling from patients with proteinuric (> 30 mg/L albumin in urine) renal disease.     

基于NMR的代谢组学方法对硝酸钕急性生物效应的研究

采用现代核磁共振和模式识别技术,分析了腹腔注射给药2、10和50mg/kg体重剂量硝酸钕48h内Wistar大鼠尿液和血清的核磁共振氢谱。由尿液及血清中内源性代谢物如柠檬酸、肌酸酐、N-氧三甲胺、氨基酸、乳酸、琥珀酸、牛磺酸及葡萄糖等物种的浓度变化,结合大鼠血清指标和肝、肾组织切片图研究了轻稀土化合物Nd(NO3)3在大鼠体内的急性生物效应。结果表明,3个剂量的Nd(NO3)3主要对大鼠肝脏造成了不同程度的损伤,而且随着剂量的升高渐趋严重。同时,Nd(NO3)3也对肾脏的特定部位(肾乳头、肾小管)造成了损害。     

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。     

Analysis of human urine metabolites using SPE and NMR spectroscopy

Nuclear magnetic resonance (NMR) spectroscopic analysis of metabonome/metabolome has widespread applications in biomedical science researches. However, most of NMR resonances for urinary metabolites remain to be fully assigned. In the present study, human urine samples from two healthy volunteers were pre-treated with C18 solid-phase extraction and the resultant 5 sub-fractions were subjected to one- and two-dimensional NMR studies, including 1H J-Resolved, 1H-1H COSY, 1H-1H TOCSY, 1H-13C HSQC, and HMBC 2D NMR. More than 70 low molecular weight metabolites were identified, and complete assignments of 1H and 13C resonances including many complex coupled spin systems were obtained.     

给药硝酸镨后大鼠尿液和血清的核磁共振代谢组学研究

采用基于核磁共振的代谢组学方法,分析了腹腔注射给药2,10和50 mg/kg体重剂量硝酸镨(Pr(NO3)3) 168 h内Wistar大鼠尿液和血清的核磁共振氢谱.由尿液及血清中内源性代谢物如柠檬酸、琥珀酸、α-酮戊二酸、肌酸酐、N-氧三甲胺、氨基酸、乳酸、牛磺酸及葡萄糖等的浓度变化,并结合大鼠血清指标研究了轻稀土化合物Pr(NO3)3在大鼠体内的急性生物效应.结果表明,Pr(NO3)3急性毒性的靶向器官为肝脏和肾脏,但以肝脏为主,且呈现明显的剂量-反应关系.低、中剂量组的Pr(NO3)3会通过改变大鼠体内酶代谢而造成肝脏线粒体中的能量代谢(脂肪、糖代谢)紊乱;同时,Pr(NO3)3还会影响肾脏的正常功能,改变肾脏中渗透质的平衡,影响肾脏对氨基酸的重吸收和利用.     

A metabonomic investigation of hepatotoxicity using diffusion-edited 1H NMR spectroscopy of blood serum

It has been shown extensively, that chemometric investigations of 1H NMR spectra of rat urine taken from animals dosed with model toxins produce characteristic patterns of metabolic responses and that this permits the identification of biomarkers of toxic response and regeneration. To date, metabonomic methods have been mainly optimised for urine which contains mainly low molecular weight moieties, and thus a conventional 1-dimensional 1H NMR pulse sequence is an efficient means of obtaining information-rich data. In the case of biofluids such as blood plasma or serum, which contain a wide range of macromolecules the resonances of which can overlap with peaks from small molecule metabolites, the information giving rise to sample classification can be concealed in a conventional NMR spectrum andthis presents a different analytical challenge in terms of chemometric analysis of spectral profiles. Here, the use of other types of NMR data have been investigated and it is shown that by using spectra where the peak intensities are edited according to their molecular diffusion coefficients, it is possible to improve differentiation of control animals and those treated with the model hepatotoxin, alpha-naphthylisothiocyanate (ANIT). By using diffusion-edited spectroscopy, plasma lipid moieties are less attenuated than those from small endogenous metabolites and thus the toxin-induced changes to the lipoprotein profiles are more easily detectable.     

SPE–NMR metabolite sub-profiling of urine

NMR-based metabolite profiling of urine is a fast and reproducible method for detection of numerous metabolites with diverse chemical properties. However, signal overlap in the (1)H NMR profiles of human urine may hamper quantification and identification of metabolites. Therefore, a new method has been developed using automated solid-phase extraction (SPE) combined with NMR metabolite profiling. SPE-NMR of urine resulted in three fractions with complementary and reproducible sub-profiles. The sub-profile from the wash fraction (100?% water) contained polar metabolites; that from the first eluted fraction (10?% methanol-90?% water) semi-polar metabolites; and that from the second eluted fraction (100?% methanol) aromatic metabolites. The method was validated by analysis of urine samples collected from a crossover human nutritional intervention trial in which healthy volunteers consumed capsules containing a polyphenol-rich mixture of red wine and grape juice extract (WGM), the same polyphenol mixture dissolved in a soy drink (WGM_Soy), or a placebo (PLA), over a period of five days. Consumption of WGM clearly increased urinary excretion of 4-hydroxyhippuric acid, hippuric acid, 3-hydroxyphenylacetic acid, homovanillic acid, and 3-(3-hydroxyphenyl)-3-hydroxypropionic acid. However, there was no difference between the excreted amounts of these metabolites after consumption of WGM or WGM_Soy, indicating that the soy drink is a suitable carrier for WGM polyphenols. Interestingly, WGM_Soy induced a significant increase in excretion of cis-aconitate compared with WGM and PLA, suggesting a higher demand on the tricarboxylic acid cycle. In conclusion, SPE-NMR metabolite sub-profiling is a reliable and improved method for quantification and identification of metabolites in urine to discover dietary effects and markers of phytochemical exposure.