中药苍耳子的毒性物质基础及中毒机制研究
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摘要
苍耳子(Fructus Xanthii),为菊科植物苍耳Xanthium sibirium Patr.的干燥成熟带总苞的果实,收载于历版中国药典,在我国资源丰富,为历代治疗鼻病及头痛的要药。然而,古代文献记载和现代临床应用都发现苍耳对人体具有一定的毒性作用,毒性一直制约着其临床应用。
关于苍耳子毒性的隐患始终未能排除,毒性成分及其作用机理未能阐明。因此,本课题首先采用小鼠急性毒性实验来明确苍耳子毒性损伤的靶器官主要为肝脏,采用体外肝细胞实验筛选毒性成分,并对目标成分进行整体动物毒性评价,在此基础上利用代谢组学技术,探讨了苍耳子的毒理学作用机制,为苍耳子的合理应用提供科学依据。本课题主要涵盖以下几方面内容:
1.苍耳子毒性物质基础研究
通过小鼠急性毒性跟踪筛选,以半数致死量LD50为指标,确定了苍耳子的毒性部位为水提取物经AB-8大孔树脂柱层析的水洗脱部位(D1),并对此部位的化学成分进行了系统的研究,从中分离出7个化合物,分别为贝壳杉烯苷类、酚酸类和含硫杂原子类化合物。
2.苍耳子毒性部位的毒性评价
采用小鼠急性毒性实验,通过病理组织学观察明确苍耳子毒性损伤的靶器官主要为肝脏,小鼠血清生化指标和肝组织匀浆生化指标检测结果表明,小鼠发生了严重的脂质过氧化损伤。本实验结果提示苍耳子的中毒原因可能是自由基攻击细胞膜中的不饱和脂肪酸,发生了脂质过氧化反应,从而导致肝细胞膜通透性的改变。在此基础上评价了苍耳子的慢性肝毒性,结果表明在长期连续重复多次给药的情况下,小鼠出现了较为严重的肝毒性反应,动物的小叶周围脂肪变性及溶解性坏死,有较多量的淋巴、浆细胞等炎细胞浸润,并伴有较明显的纤维组织增生。由于慢性鼻炎、鼻窦炎等疾病需长期用药,剂量使用不当可引起慢性蓄积中毒,导致肝功能损害。所以长期、大剂量服用苍耳子对机体的不良影响应引起足够的重视。
3.毒性成分的筛选和毒性评价
采用人正常肝细胞株L-02和正常大鼠肝细胞株BRL作为体外细胞毒性的试验模型,比较研究苍耳子主要成分的细胞毒性。结果表明苍术苷、羧基苍术苷和4’-去磺基苍术苷随着浓度逐渐升高,抑制作用逐渐增强,浓度为10-4 mol/L时能够明显抑制对两种细胞的增殖作用。苍术苷等造成了不同程度的细胞皱缩现象,细胞间隙扩大,随着给药浓度增加,细胞出现空泡化,死细胞明显增多。苍术苷等引起LDH漏出增加,细胞通透性增加,具有一定的细胞毒作用。并采用整体动物实验对目标成分苍术苷进行了毒性评价,结果表明苍术苷高剂量组对小鼠的肝脏造成了较严重的损伤,且中毒机理可能与苍耳子毒性部位相似,证明了苍耳子中的主要毒性成分为贝壳杉烯苷类化合物,与体外实验结果相符。
4.利用代谢组学技术探讨苍耳子的毒性作用机制
利用1H NMR技术研究苍耳子处理后大鼠尿液和血浆中内源性代谢产物谱的变化,研究发现尿液中乳酸盐、葡萄糖、马尿酸盐、醋酸盐、三甲胺N氧化物、色氨酸、二甲胺、肌醇、柠檬酸盐、N甲基烟酸盐和酪氨酸明显升高,而酮戊二酸、肌酐、异戊酸盐、琥珀酸盐、组氨酸、丁酸盐、二甲基甘氨酸、N-乙酰基-半胱氨酸、牛磺酸和赖氨酸含量下降。血浆中异戊酸盐、乳酸盐、缬氨酸、谷氨酸盐、高密度脂蛋白、乙酸盐、肌酐、丙酮和尿素升高,而甘氨胆碱和葡萄糖明显下降。柠檬酸盐是三羧酸循环的中间产物,主要在肝细胞线粒体中进行代谢;琥珀酸盐的含量的降低是由于降低脂肪酸分解代谢。这些成分的变化影响肝细胞线粒体的能量代谢;烟酸盐及甲基烟酰胺均为色氨酸NAD+通路的产物,色氨酸代谢产物的升高伴随肝脏中脂质过氧化酶的生成减少,影响局部氧化还原平衡。肌醇,乳酸的浓度升高,是体内缺氧代谢的结果。所以苍耳子的毒性主要与其影响了肝细胞能量代谢有关。尿液中的柠檬酸盐、马尿酸盐、色氨酸等可以作为苍耳子的主要生物标志物研究。
综上,苍耳子毒性作用靶器官主要是肝脏,贝壳杉烯苷类化合物是主要的毒性成分,其毒性作用与苍耳子引起动物肝脏的脂质过氧化损伤和影响肝细胞能量代谢机理有关。本课题将代谢组学与传统毒理学研究方法相结合,初步揭示了中药苍耳子毒性作用的肝损伤机制。我们的研究显示,用代谢组学方法揭示的生物化学变化很容易与传统手段的测定结果相联系,更容易发现药物作用的生物化学物质基础和作用机制,因此,药物毒性作用机制的代谢组学研究必将在成分复杂的中药体系中发挥更大的作用。
苍耳子的临床应用十分广泛,正常情况下使用苍耳子是安全的,但长期、大剂量使用会出现毒性反应,因此在临床上长期用药不要超过3-9 g,肝损伤患者应用时需要调整剂量,长期应用时要注意监测其肝功能。其他含贝壳杉烯苷类中药可能也有类似的毒性毒理机制,因此,应该建立质量标准,控制毒性成份的含量,加强对此类中药的不良反应监测。随着代谢组学技术的发展,此项技术将会越来越多的用于其他中草药毒性毒理的研究,对于实现中药现代化具有重要的意义。
Fructus Xanthii named Cang-Er-Zi is the ripe fruits with involucre of Xanthium sibirium Patr. It is a traditional Chinese medicine that is used in curing nasal diseases and headache according to the Chinese Pharmacopoeia. Fructus Xanthii is in great need in clinic. However, utilization of Fructus Xanthii is limited by its toxicity. Fructus Xanthii is toxicant,and it maybe result in poisoning when used excessively,without preparation or in inappropriate preparation.
Recently studies have demonstrated that Fructus Xanthii may have the potential of hepatotoxicity. However, so far there is little studies concening the toxic mechanisms and the toxicologic assessment of its essential components. So, there are no identified conclusions about its toxicity. In this study, in vivo and in vitro experiments have been applied to explore the toxicity and the target organs of Fructus Xanthii, and compared the toxicity of its essential components. After then, the toxic mechanisms are elucidated based on metabonomic technologies in order to offer some toxicological information for its safely clinical usage. The main contents of our study are as following three aspects:
1 Study on toxic components of Fructus Xanthii
Toxicity response of different extractions from Fructus Xanthii was compared using acute toxicity experiment LD50 and MTD in mice. Toxic fraction(D1) was determined. Bioactivity guided isolation of toxic fractions and compounds were undertaken by various chromatographic methods. 7 known compounds were isolated from the toxic fraction. The major types of the isolated compounds were phenolic acids, kaurene glucosides and heterocycles.
2 Evaluation of toxic fractions of Fructus Xanthii
The results of level of serum transaminase, biochemical parameters of liver tissue and histopathlogy showed that the main toxic target organ of Fructus Xanthii is liver using acute toxicity experiment. It demonstrated that Fructus Xanthii induce hepatotoxicity in mice by way of its induction of oxidative stress as lipid peroxidation in liver, which merited further studies. Then, the chronic liver toxicity experiment was also determined. It deserves attention that long-term and high-dosage administration of Fructus Xanthii can damage the liver to varied degrees.
3 Evaluation of toxic components of Fructus Xanthii
Comparative toxicity studies of its essential components indicated that atractyloside, carbxyatractyloside and 4’-desulphate-atractyloside are main toxic constituents using cell test. It was found that the three kaurene glycosides can strongly inhibite human normal liver L-02 and rat normal liver BRL cells growth. Then, the toxicity of atractyloside was determined in toxicity experiment in mice. The results demonstrated that the toxic constituents of Fructus Xanthii are kaurene glucosides.
4 Investigation of toxic methanism of Fructus Xanthii using integrated metabonomic technology
A integrated metabonomic study with high-resolution 1H NMR spectroscopy has been applied to investigate the biochemical composition of urine and plasma obtained from Fructus Xanthii treated rats. It was found that the level ofβ-HB, lactate, valine, Glutamate, Creatinine, acetone, lactate and allantoin was elevated,and the levels of GPC and glucose were decreased in blood plasma. The level of lactate, hippurate, acetate, TMAO, tryptophane, MA, myoinositol, citrate, N-methy-nicotinamide and tyrosine increased in urine,whileα-ketoglutarate, Creatinine, 2-hydroxy-isovalerate, succinate, histidine, 3-hydroxy butyrate, DMG, NAC, taurine and lysine decreased significantly. The increased level of citrate, N-methy-nicotinamide and tryptophane is the typical biomarker of induction of oxidative stress as lipid peroxidation in liver of Fructus Xanthii.
Above all, the main toxic target organ of Fructus Xanthii is liver. Kaurene glycosides are main toxic constituents. From these results, we can infer that Fructus Xanthii induce hepatotoxicity in mice by way of its lipid peroxidation and energy metabolismin. Combination with biochemistry and histological research,we used metabonomic analysis of urines and serum samples to visualize significant alterations in metabolite expression patterns as a result of induced metabolic responses. The study of these metabolite alterations also allowed several major metabolic pathways.
Fructus Xanthii is widely and safely used in Chinese medicines usually, while the toxic effects will appear when it was used in large dose and for a long time. There for, when it is used for a long time, the dose will not exceed 3-9 g. When the patients have liver injury, the dose must be modified, and functions of kidney and liver should be monitored. Other herb containing kaurene glycosides may have similar toxic effects. Quality criteria should be established to control the content of toxic substance acordingly. With the development of metabonomics,this novel technology might provide additional discrimination in the toxicological effects of other herb in the future, which will contributes to the modemlization of traditional Chinese medicines.
关于苍耳子毒性的隐患始终未能排除,毒性成分及其作用机理未能阐明。因此,本课题首先采用小鼠急性毒性实验来明确苍耳子毒性损伤的靶器官主要为肝脏,采用体外肝细胞实验筛选毒性成分,并对目标成分进行整体动物毒性评价,在此基础上利用代谢组学技术,探讨了苍耳子的毒理学作用机制,为苍耳子的合理应用提供科学依据。本课题主要涵盖以下几方面内容:
1.苍耳子毒性物质基础研究
通过小鼠急性毒性跟踪筛选,以半数致死量LD50为指标,确定了苍耳子的毒性部位为水提取物经AB-8大孔树脂柱层析的水洗脱部位(D1),并对此部位的化学成分进行了系统的研究,从中分离出7个化合物,分别为贝壳杉烯苷类、酚酸类和含硫杂原子类化合物。
2.苍耳子毒性部位的毒性评价
采用小鼠急性毒性实验,通过病理组织学观察明确苍耳子毒性损伤的靶器官主要为肝脏,小鼠血清生化指标和肝组织匀浆生化指标检测结果表明,小鼠发生了严重的脂质过氧化损伤。本实验结果提示苍耳子的中毒原因可能是自由基攻击细胞膜中的不饱和脂肪酸,发生了脂质过氧化反应,从而导致肝细胞膜通透性的改变。在此基础上评价了苍耳子的慢性肝毒性,结果表明在长期连续重复多次给药的情况下,小鼠出现了较为严重的肝毒性反应,动物的小叶周围脂肪变性及溶解性坏死,有较多量的淋巴、浆细胞等炎细胞浸润,并伴有较明显的纤维组织增生。由于慢性鼻炎、鼻窦炎等疾病需长期用药,剂量使用不当可引起慢性蓄积中毒,导致肝功能损害。所以长期、大剂量服用苍耳子对机体的不良影响应引起足够的重视。
3.毒性成分的筛选和毒性评价
采用人正常肝细胞株L-02和正常大鼠肝细胞株BRL作为体外细胞毒性的试验模型,比较研究苍耳子主要成分的细胞毒性。结果表明苍术苷、羧基苍术苷和4’-去磺基苍术苷随着浓度逐渐升高,抑制作用逐渐增强,浓度为10-4 mol/L时能够明显抑制对两种细胞的增殖作用。苍术苷等造成了不同程度的细胞皱缩现象,细胞间隙扩大,随着给药浓度增加,细胞出现空泡化,死细胞明显增多。苍术苷等引起LDH漏出增加,细胞通透性增加,具有一定的细胞毒作用。并采用整体动物实验对目标成分苍术苷进行了毒性评价,结果表明苍术苷高剂量组对小鼠的肝脏造成了较严重的损伤,且中毒机理可能与苍耳子毒性部位相似,证明了苍耳子中的主要毒性成分为贝壳杉烯苷类化合物,与体外实验结果相符。
4.利用代谢组学技术探讨苍耳子的毒性作用机制
利用1H NMR技术研究苍耳子处理后大鼠尿液和血浆中内源性代谢产物谱的变化,研究发现尿液中乳酸盐、葡萄糖、马尿酸盐、醋酸盐、三甲胺N氧化物、色氨酸、二甲胺、肌醇、柠檬酸盐、N甲基烟酸盐和酪氨酸明显升高,而酮戊二酸、肌酐、异戊酸盐、琥珀酸盐、组氨酸、丁酸盐、二甲基甘氨酸、N-乙酰基-半胱氨酸、牛磺酸和赖氨酸含量下降。血浆中异戊酸盐、乳酸盐、缬氨酸、谷氨酸盐、高密度脂蛋白、乙酸盐、肌酐、丙酮和尿素升高,而甘氨胆碱和葡萄糖明显下降。柠檬酸盐是三羧酸循环的中间产物,主要在肝细胞线粒体中进行代谢;琥珀酸盐的含量的降低是由于降低脂肪酸分解代谢。这些成分的变化影响肝细胞线粒体的能量代谢;烟酸盐及甲基烟酰胺均为色氨酸NAD+通路的产物,色氨酸代谢产物的升高伴随肝脏中脂质过氧化酶的生成减少,影响局部氧化还原平衡。肌醇,乳酸的浓度升高,是体内缺氧代谢的结果。所以苍耳子的毒性主要与其影响了肝细胞能量代谢有关。尿液中的柠檬酸盐、马尿酸盐、色氨酸等可以作为苍耳子的主要生物标志物研究。
综上,苍耳子毒性作用靶器官主要是肝脏,贝壳杉烯苷类化合物是主要的毒性成分,其毒性作用与苍耳子引起动物肝脏的脂质过氧化损伤和影响肝细胞能量代谢机理有关。本课题将代谢组学与传统毒理学研究方法相结合,初步揭示了中药苍耳子毒性作用的肝损伤机制。我们的研究显示,用代谢组学方法揭示的生物化学变化很容易与传统手段的测定结果相联系,更容易发现药物作用的生物化学物质基础和作用机制,因此,药物毒性作用机制的代谢组学研究必将在成分复杂的中药体系中发挥更大的作用。
苍耳子的临床应用十分广泛,正常情况下使用苍耳子是安全的,但长期、大剂量使用会出现毒性反应,因此在临床上长期用药不要超过3-9 g,肝损伤患者应用时需要调整剂量,长期应用时要注意监测其肝功能。其他含贝壳杉烯苷类中药可能也有类似的毒性毒理机制,因此,应该建立质量标准,控制毒性成份的含量,加强对此类中药的不良反应监测。随着代谢组学技术的发展,此项技术将会越来越多的用于其他中草药毒性毒理的研究,对于实现中药现代化具有重要的意义。
Fructus Xanthii named Cang-Er-Zi is the ripe fruits with involucre of Xanthium sibirium Patr. It is a traditional Chinese medicine that is used in curing nasal diseases and headache according to the Chinese Pharmacopoeia. Fructus Xanthii is in great need in clinic. However, utilization of Fructus Xanthii is limited by its toxicity. Fructus Xanthii is toxicant,and it maybe result in poisoning when used excessively,without preparation or in inappropriate preparation.
Recently studies have demonstrated that Fructus Xanthii may have the potential of hepatotoxicity. However, so far there is little studies concening the toxic mechanisms and the toxicologic assessment of its essential components. So, there are no identified conclusions about its toxicity. In this study, in vivo and in vitro experiments have been applied to explore the toxicity and the target organs of Fructus Xanthii, and compared the toxicity of its essential components. After then, the toxic mechanisms are elucidated based on metabonomic technologies in order to offer some toxicological information for its safely clinical usage. The main contents of our study are as following three aspects:
1 Study on toxic components of Fructus Xanthii
Toxicity response of different extractions from Fructus Xanthii was compared using acute toxicity experiment LD50 and MTD in mice. Toxic fraction(D1) was determined. Bioactivity guided isolation of toxic fractions and compounds were undertaken by various chromatographic methods. 7 known compounds were isolated from the toxic fraction. The major types of the isolated compounds were phenolic acids, kaurene glucosides and heterocycles.
2 Evaluation of toxic fractions of Fructus Xanthii
The results of level of serum transaminase, biochemical parameters of liver tissue and histopathlogy showed that the main toxic target organ of Fructus Xanthii is liver using acute toxicity experiment. It demonstrated that Fructus Xanthii induce hepatotoxicity in mice by way of its induction of oxidative stress as lipid peroxidation in liver, which merited further studies. Then, the chronic liver toxicity experiment was also determined. It deserves attention that long-term and high-dosage administration of Fructus Xanthii can damage the liver to varied degrees.
3 Evaluation of toxic components of Fructus Xanthii
Comparative toxicity studies of its essential components indicated that atractyloside, carbxyatractyloside and 4’-desulphate-atractyloside are main toxic constituents using cell test. It was found that the three kaurene glycosides can strongly inhibite human normal liver L-02 and rat normal liver BRL cells growth. Then, the toxicity of atractyloside was determined in toxicity experiment in mice. The results demonstrated that the toxic constituents of Fructus Xanthii are kaurene glucosides.
4 Investigation of toxic methanism of Fructus Xanthii using integrated metabonomic technology
A integrated metabonomic study with high-resolution 1H NMR spectroscopy has been applied to investigate the biochemical composition of urine and plasma obtained from Fructus Xanthii treated rats. It was found that the level ofβ-HB, lactate, valine, Glutamate, Creatinine, acetone, lactate and allantoin was elevated,and the levels of GPC and glucose were decreased in blood plasma. The level of lactate, hippurate, acetate, TMAO, tryptophane, MA, myoinositol, citrate, N-methy-nicotinamide and tyrosine increased in urine,whileα-ketoglutarate, Creatinine, 2-hydroxy-isovalerate, succinate, histidine, 3-hydroxy butyrate, DMG, NAC, taurine and lysine decreased significantly. The increased level of citrate, N-methy-nicotinamide and tryptophane is the typical biomarker of induction of oxidative stress as lipid peroxidation in liver of Fructus Xanthii.
Above all, the main toxic target organ of Fructus Xanthii is liver. Kaurene glycosides are main toxic constituents. From these results, we can infer that Fructus Xanthii induce hepatotoxicity in mice by way of its lipid peroxidation and energy metabolismin. Combination with biochemistry and histological research,we used metabonomic analysis of urines and serum samples to visualize significant alterations in metabolite expression patterns as a result of induced metabolic responses. The study of these metabolite alterations also allowed several major metabolic pathways.
Fructus Xanthii is widely and safely used in Chinese medicines usually, while the toxic effects will appear when it was used in large dose and for a long time. There for, when it is used for a long time, the dose will not exceed 3-9 g. When the patients have liver injury, the dose must be modified, and functions of kidney and liver should be monitored. Other herb containing kaurene glycosides may have similar toxic effects. Quality criteria should be established to control the content of toxic substance acordingly. With the development of metabonomics,this novel technology might provide additional discrimination in the toxicological effects of other herb in the future, which will contributes to the modemlization of traditional Chinese medicines.
引文
[1]陈一凡,蔡皓东.中药引起肝损害的调查分析.药物不良反应杂志,l999,1(1):27-32.
[2]邓培嫒,傅琪.116例药源性肝损害分析.药物流行病学杂志,1998,7(3):144-145.
[3]徐列明.对中草药引起肝损害的认识及防治.现代医药卫生,2005,21(4):379-380.
[4]陈勇,程明,刘春霞.肝脏毒性中药的研究现状与展望.中草药,2004,35(11):1315-1317.
[5]傅肖岩,劳绍贤.重视中药所致肝损害.中药新药与临床药理,2003,14(2):130-133.
[6]胡锡琴.对中草药引起药源性肝损害的分析.陕西中医,2002,23(9):837-838.
[7]卢晓露,张煜,赵肃清.具肝毒性天然药物的研究进展及防治策略.现代食品与药品杂志,2007,17(1):11-15.
[8]中华人民共和国药典委员会.中华人民共和国药典[S].一部.北京:化学工业出版社,2005:111-112.
[9]李全林,朱桂芳.复方苍耳子油剂治疗鼻窦炎98例.陕西中医,2009,30(12):1644.
[10]骆桂秋,顾析玲,宋宪华.苍耳子汤配合西药治疗小儿急性鼻窦炎73例—附单纯西药治疗62例对照.浙江中医杂志,2005,40(7):289-289.
[11]李红,周谋.苍耳子及复方制剂的药理作用和临床研究进展.山西医科大学学报,2004,35(3):313-314.
[12]华刚,孟静.玉屏风散合苍耳子散加减治疗过敏性36例.河南中医,2005,25(3):65-65.
[13]侯自强.连蒲苍耳子散联合药物冲洗治疗慢性鼻窦炎138例疗效观察.中医药导报,2009,15(8):52-54
[14]韩婷.苍耳子的生物活性成分及品质评价(博士论文).上海:第二军医大学,2006.
[15]韩婷,秦路平,郑汉臣.苍耳及其同属药用植物研究进展.解放军药学学报,2003,19(2):122-125.
[16]Lu-ping Qin,Ting Han,Huiliang Li,Qiaoyan Zhang,et al. A new thiazinedione from Xanthium strumarium. Fitoterapia, 2006,77: 245-246.
[17]Ting Han,Huiliang Li,Qiaoyan Zhang,et al. New thiazinediones and other components from Xanthium strumarium L.. Chem Nat Compd, 2006,42(5): 567-570.
[18]张晓琦.苍耳属中倍半萜内酯的研究进展.中草药,2001,32(10):951-953.
[19]梅全喜,毕焕新.现代中药药理手册.北京:中国中医药出版社,1998,41-43.
[20]宋振玉,张凌云,谢明智,等.苍耳子有毒成分及药理作用.药学学报,1962,9(11):678-683.
[21]何芙莉,闫姝,张小波.苍耳子小鼠最大耐受量实验观察.中国中西医结合外科杂志, 1998,4(2):107-108.
[22]MacLeod JK,Moeller PD,Franke FP. Two toxic glycosides from the burrs of Xanthium pungens. J Nat Prod, 1990, 53(3): 451.
[23]王素贤,任丽娟,孙泽人.蒙古苍耳种仁中的有毒成分.中草药,1983,14(12):1.
[24]苏亚,翁维良,胡瑾.临床中药学.郑州:河南科技出版社,1998,214-215.
[25]张学梅,张重华.苍耳子中毒及毒性研究进展.中西医结合学报.2003,1(1):71-73.
[26]郭汉林.急性鲜苍耳子中毒12例报告.临床内科杂志,1989,6(2):29.
[27]周加权.以多器官损害为表现的苍耳子中毒1例报告.中国社区医师,2005,21(5):47.
[28]赵胜乾,吴敏.苍耳子中毒致急性肾功能衰竭及肝损伤1例.实用诊断与治疗杂志,2004,18(6):514.
[29]梁晓燕,朱会友,王建丽.小剂量苍耳子中毒1例.实用中医药杂志,1998,14(10):45-46.
[30]丁雅芳.简述中药毒副反应.湖北中医杂志,1985,4:43.
[31]纪绍先,张国栋.急性苍耳子中毒12例报告.吉林医科大学学报,1962,1(1):75-78.
[32]杜青云,潘东林.73例急性中药中毒分析.药学实践杂志,1995,13(5):277-279.
[33]王希海,黄光照.药用植物引起中毒性肝病时的病理变化.中西医结合肝病杂志,1996, 6(4):48-50.
[34]朱天忠.试论中草药的肝毒作用.中国法医学杂志,1986,1(1):56.
[35]王文英.部分中药对肾脏毒副作用及防治.时珍国医国药,2000,11(4):347-348.
[36]赵宝玉,王建华,史克诚.动物毒理学.北京:中国农业出版社,2001,349-351.
[37]钱培忠.有毒中药大辞典.天津:天津科技出版公司,1995,258-260.
[38]姜萧,隋进江.浅谈中药苍耳子毒性及中毒的抢救方法.黑龙江中医药,2000,20(4):63.
[39]李涓,高天,谢子清,唐熠.苍耳子不同提取物的毒性比较实验.时珍国医国药,2005,16(6)484-488.
[40]Nishimura H,Nonaka GI,Nishioka I. Seven quinic acid gallates from Quercus stenophylla. Phytochemistry,1984,23 (11):2621-2623.
[41]Morishita H, et a1.Chromatographic separation and identification of naturally occurring choirogenic acids by 1H NMR spectroscopy and mass spectrometry. J Chromatogr 1984,315:253.
[42]郑晓珂,李钦,冯卫生.冬凌草中酚酸类化学成分研究.中国药学杂志,2004,39(5):335-336.
[43]Ma YT, Huang MC, Hsu FL, et al. Thiazinedion from Xanthium strumarium. Phytochemistry, 1998, 48(6): 1083-1085.
[44]Mahmoud,Ahmed A, Al-Shihry, Shar S, et al. A new heterocyclic glucoside from the fruits of Xanthium pungens.Natural Product Research,2005,19(6): 585-589.
[45]Sonia P,Cosimo P,Nunziatina DT,et al. Sesquiterpene and diterpene glycosides from Xanthium spinosum.Phytochemistry,1996,41(5):1357-60.
[46]翁维良.临床中药不良反应.中药新药与临床药理,1996,7(2):4.
[47]雷树勇,黄承乐,陆爱权.11种生化项目在肝脏疾病中的变化分析.2002,14(1):59-61.
[48]刘娓娓,顾文君,沈锡中.血清生化指标对慢性乙型肝炎肝纤维化及炎症程度的诊断价值.中国临床医学,2005,12(1):65-66.
[49]周加权.以多器官损害为表现的苍耳子中毒1例报告.中国社区医师,2005,21(5):47.
[50]叶秀珍,朱德厚,沈鼎武.体外连续培养的成人肝细胞的超显微结构.实验生物学报, 1980,13:3612-3641.
[51]郭竹英,矫强,徐芒华,王世婷.脂多糖对人正常肝细胞株L02损伤的实验研究.医学分子生物学杂志,2008,5(4):323-327.
[52]陈莹,季莉莉,马鸿雁,刘天瑜,王峥涛.吡咯里西啶生物碱Jacoline对人正常肝L-02细胞内谷胱甘肽含量及还原酶活性的影响.上海中医药大学学报.2008,22(2):35-39.
[53]孟雪莲,曾晓非,孙祖越,裴晓丹.丹参致大鼠肝细胞脂质过氧化损伤的实验研究.辽宁大学学报,2005,32(1):31-34.
[54]周成江,周立社,和彦苓,杨艳芳.蒙药童格勒格四种粗提物对大鼠正常肝细胞BRL株增殖的影响.包头医学院学报,2008,24(4):348-349.
[55]陈国勇,曲青山,孙建军,钱建民.转染血红素氧化酶21基因大鼠肝细胞系BRL体外抗凋亡作用.郑州大学学报(医学版),2006,41(4):712-714.
[56]马巧玉,王宇明,郝飞.LDH检测BALB/c小鼠CTL活性的初步研究.第三军医大学学报, 2003,25(4):298-300.
[57]王甫厚,陈绍先,郭彩云,等.LDH释放法测NK细胞毒的方法学研究.中国免疫学杂志, 1990,6(2):115.
[58]张蕾,李芳秋,张士新,黄海嵘,李德闵.LDH释放法检测肿瘤患者PBMC和CIK细胞的细胞毒活性.现代检验医学杂志,2008,23(6):27-29.
[59]Cole RJ,Stuart BP,Lansden JA,Cox RH.Isolation and redefinition of the toxic agent fromcocklebur (Xanthium strumarium). J Agric Food Chem,1980,28: 1330-1332.
[60]Georgiou M,Sianidon L,Hatziz T,Papadatos J,Konselini SA. Hepatotoxicity due to Atractylis gummifera L.Clin Toxicol 1988,26:487-493.
[61]Larrey D.Hepatotoxicity of herbal remedies.J Hepatol 1997,26:47-54.
[62]Mokhobo KP.Herb use and necro-degenerative hepatitis.S Afr Med J 1976,50:1096-9.
[63]Luciani S,Carpenedo F,Tarjan EM. Effects of atractyloside and carboxyatracty-loside in the whole animal. In Atractyloside: Chemistry, Biochemistry and Toxicology1978,109.
[64]Obatomi DK, Bach PH.Biochemistry and Toxicology of the Diterpenoid Glycoside Atractyloside.Food and Chemical Toxicology 1998,36:335-346.
[65]Sinclair AJ, et al. NMR-based metabolomic analysis of cerebrospinal fluid and serum in neurological diseases - a diagnostic tool.NMR Biomed, 2010,23(2):123-132.
[66]Lee JE, et al.Evidence of vintage effects on grape wines using 1H NMR-based metabolomic study.Anal Chim Acta,2009.648(1):71-76.
[67]Lee IJ, et al. NMR metabolomic analysis of caco-2 cell differentiation. J Proteome Res, 2009,8(8):4104-4108.
[68]Ko BK, et al. Metabolomic insight into soy sauce through 1H NMR spectroscopy. J Agric Food Chem,2009,57(15):6862-6870.
[69]Hirakawa K, et al. Experimental estimation of postmortem interval using multivariate analysis of proton NMR metabolomic data. Leg Med (Tokyo),2009,11(1):282-285.
[70]Nicholas PC, et al. 1H NMR-based metabolomic analysis of liver,serum,and brain following ethanol administration in rats.Chem Res Toxicol,2008,21(2):408-420.
[71]Kruger NJ, Troncoso-Ponce MA, Ratcliffe RG. 1H NMR metabolite fingerprinting and metabolomic analysis of perchloric acid extracts from plant tissues. Nat Protoc,2008,3(6):1001-1012.
[72]Poulding S, et al. Removal of t(1) noise from metabolomic 2D(1)H-(13)C HSQC NMR spectra by Correlated Trace Denoising.J Magn Reson, 2007.189(2): 190-199.
[73]Gu H, et al. 1H NMR study of the effects of sample contamination in the metabolomic analysis of mouse urine.J Pharm Biomed Anal,2007,45(1):134-140.
[74]Cho IH, Kim YS, Choi HK. Metabolomic discrimination of different grades of pine mushroom (Tricholoma matsutake Sing.) using 1H NMR spectrometry and multivariate dataanalysis.J Pharm Biomed Anal,2007,43(3):900-904.
[75]Zhou J, et al.1H NMR-based metabonomic and pattern recognition analysis for detection of oral squamous cell carcinoma.Clin Chim Acta,2009,401(1):8-13.
[76]Xiao C, et al. Revealing the metabonomic variation of rosemary extracts using 1H NMR spectroscopy and multivariate data analysis.J Agric Food Chem,2008,56(21): 142-153.
[77]Craig A, et al.Scaling and normalization effects in NMR spectroscopic metabonomic data sets.Anal Chem,2006,78(7):2262-2267.
[78]Webb-Robertson BJ, et al. A study of spectral integration and normalization in NMR-based metabonomic analyses.J Pharm Biomed Anal,2005,39(3-4):830-836.
[79]Bundy JG, et al. A NMR-based metabonomic approach to the investigation of coelomic fluid biochemistry in earthworms under toxic stress.FEBS Lett,2001,500(1-2):31-35.
[80]Chan ECY, et al. Metabolic Profiling of Human Colorectal Cancer Using High–Resolution Magic Angle Spinning Nuclear Magnetic Resonance (HR-MAS NMR) Spectroscopy and Gas Chromatography Mass Spectrometry (GC/MS).J Proteome Res,2009,8(1):352-361.
[81]Yamamoto T, Horii I, Yoshida T.Integrated NMR-based metabonomic investigation of early metabolic effects of ethylene glycol monomethyl ether (EGME) on male reproductive organs in rats.J Toxicol Sci,2007,32(5):515-528.
[82]Psihogios NG, et al.Evaluation of tubulointerstitial lesions severity in patients with glomeru-onephritides:an NMR-based metabonomic study. J Proteome Res,2007, 6(9):3760-3770.
[83]Bertram HC,et al. Effect of magnetic field strength on NMR-based metabonomic human urine data.Comparative study of 250,400,500,and 800 MHz. Anal Chem, 2007,79(18): 7110-7115.
[84]Bertram HC,et al. NMR-based metabonomic studies reveal changes in the biochemical profile of plasma and urine from pigs fed high-fibre rye bread. Br J Nutr, 2006. 95(5): 955-962.
[85]Zha W, et al.Metabonomic characterization of early atherosclerosis in hamsters with induced cholesterol. Biomarkers,2009.14(6):372-380.
[86]Yin P, et al.A metabonomic study of hepatitis B-induced liver cirrhosis and hepatocellular carcinoma by using RP-LC and HILIC coupled with mass spectrometry.MolBiosyst,2009,5(8):868-876.
[87]Sieber M, et al. Metabonomic Study of Ochratoxin A Toxicity in Rats after Repeated Administration:Phenotypic Anchoring Enhances the Ability for Biomarker Discovery.Chem Res Toxicol,2009,22(7):1221-1231.
[88]Yan S, et al. Metabonomic characterization of aging and investigation on the anti-aging effect of total flavones of Epimedium.Mol Biosyst, 2009,5(10):1204-1213.
[89]Huang X, et al.A metabonomic characterization of CCl4-induced acute liver failure using partial least square regression based on the GC/MS metabolic profiles of plasma in mice.J Chromatogr B Analyt Technol Biomed Life Sci,2008,870(2):178-185.
[90]Fardet A, et al.Whole-grain and refined wheat flours show distinct metabolic profiles in rats as assessed by a 1H NMR-based metabonomic approach. J Nutr,2007.137(4):923-929.
[91]Keun HC. Metabonomic modeling of drug toxicity. Pharmacology Therapeutics, 2006, 109(1-2):92-106.
[92]Beckonert O, et al. NMR-based metabonomic toxicity classification: hierarchical cluster analysis and k-nearest-neighbour approaches. Anal. Chim. Acta,2003.490(1-2): 3-15.
[93]Nicholls AW, et al. Metabonomic investigations into hydrazine toxicity in the rat. Chem Res Toxicol.,2001,14(8):975-987.
[94]Bollard ME, et al.Comparative metabonomics of differential hydrazine toxicity in the rat and mouse.Toxicol.Appl Pharmacol,2005,204(2):135-151.
[95]Mortishire-Smith RJ, et al.Use of metabonomics to identify impaired fatty acid metabolism as the mechanism of a drug-induced toxicity.Chem Res Toxicol,2004,17(2):165-173.
[96]Ebbels T, et al.Toxicity classification from metabonomic data using a density superposition approach:'CLOUDS'.Anal Chim Acta,2002,490(1-2):109-122.
[97]Wu H, et al. NMR spectroscopic-based metabonomic investigation on the acute biochemical effects induced by Ce(NO3)3 in rats. J Inorg Biochem,2005,99(11):2151-2160.
[98]Zhang XY, et al. NMR-based metabonomic study on the subacute toxicity of aristolochic acid in rats.Food and Chemical Toxicology,2006,44(7):1006-1014.
[99]李中港,程芳,盛卸晃,刘兆平.代谢组学在药物安全性评价中的应用进展.毒理学杂志. 2005,24(5):415-418.
[100]王全军,颜贤忠,吴纯启.Z24经口染毒大鼠尿液的核磁共振谱代谢组学研究.中国药理学与毒理学杂志,2004,18(6):465-470.
[101]王全军,颜贤忠,吴纯启.Z24经口染毒大鼠血浆的代谢组学研究.卫生毒理学杂志,2004,18(2):73-75.
[102]李伶,颜贤忠.代谢组学在毒理学中的应用进展.军事医学科学院院刊,2007,31(2): 183-186.
[103]何君,周宏灏.代谢组学及其在药理学中的进展.中国药理学通报,2006,22(11):1304-1309.
[104]Nicholson JK,Conelly J,Lindon JC,et al. Metabonomics:A Platform for studying drug toxicity and gene function.Nat Rev Drug Discov,2002,1(2):153-161.
[105]Holmes E, Nicholls AW, Lindon JC,et al. A development of a model for classification of toxin-induced lesions using lH NMR spectroscopy of urine combined with pattern recognition.NMRBiomed,l998,11(4-5):235-244.
[106]Holmes E,Nicholson JK, NiCholls AW,et al.The identification of novel Biomarkers of renal toxicity using automatic data reduction techniques and PCA of Proton NMR spectra of urine.Chemomet.lntel Lab systems,1998,44:245-255.
[107]Holmes E,Nicholls AW,Lindon JC,et al.Chemometric models for toxicity classification based on NMR spectra of biofluids.Cheml Res Toxicol,2000,13(6):471-478.
[108]Nicholson JK,lindon J,Holmes E.Metabonomics:understanding the Metabolic responses of living systems to pathophysiological stimuli via muItivariate statisticaI analysis of biological NMR spectroscopic data Xenobiotica,1999,29(11):1181-1189.
[1] Nicholson JK,Wilson ID. Opinion:understanding global systems biology:metabonomics and the continuum of metabolism.Nat Rev Drug Discov,2003,2(8):668-676
[2] Fiehn O. Combining genomics, metabolome analysis, and biochemical modelling to understand metabolic networks. Comp Funct Genom, 2001, 2: 155-168
[3] Fiehn O. Metabolomics-the link between genotypes and phenotypes.Plant Mol Biol, 2002, 48: 155-171
[4] Nicholson JK, Connelly J, Lindon JC,et al. Metabonomics: a plat-form for studying drug toxicity and gene function.Nat Rev Drug Discov,2002,1(2):153
[5] Lindon JC,Nicholson JK,Holmes E. Metabonomics: Metabolic processes studied by NMR spectroscopy of biofluids. Concepts Magn Reson,2000,12,5:289-320
[6] Lindon JC, Holmes E, Nicholson JK. Recent development and Applications of NMR-Based Metabonomics. Chinese Journal of Magnetic Resonance, 2006,23(1):101-127
[7] Plumb RS,Stumpf CL,Granger JH,et al. Use of liquid chro-matography/time-of-flight mass spectrometry and multivariate statistical analysis shows promise for the detection of drug metabolites in biological fluids.Papid Communic in Mass Spectrome,2003,17(23): 2632-2638
[8] Plumb RS,Stumpf CL,Gorenstein MV, et al. Metabonomics:the use of electrospray mass spectrometry coupled to reversed phase liquid chromatography shows potential for the screening on rat urine in drug development.Rapid Communic in MasSpectrome, 2002, 16(20): 1991-1996
[9] Beaudry F,Le Blanc JC Y,Coutu M,et al. Metabolite profiling study of propranolol in ratusing LC/MS/MS analysis.Biomed Chromatogra,1999,13(5): 363-369
[10] Hai PT, Kaskavelis L, Daykin CA, Janssen HJ. Method development in high-performance liquid chromatography for high-throughput profiling and metabonomic studies of biofluid samples. Chromatogr B2003, 789(2): 283-301
[11] Plumb RS, Stumpf CL, Gorenstein MV, Castro-Perez JM, Dear GJ, Anthony M, Sweatman BC, Connor SC, Haselden JN. Metabonomics: the use of electrospray mass spectrometry coupled to reversed-phase liquid chromatography shows potential for the screening of rat urine in drug development.Rapi Commun Mass Spec, 2002, 16: 1991-1996
[12] Bugrim A,Nikolskaya T,Nikolsky Y. Early prediction of drug metabolism and toxicity: systems biology approach and modeling.Drug DiscovT 2004,9(3): 127-135
[13] Taylor J, King RD, AltmannT, et al. Application of metabolomics to plant genotype discrimination using statistics and machine learning.Bioinformatics,2002,18(2): 241
[14] Chen MJ, Zhao LP, Jia W.Metabonomic study on the biochemical profiles of a hydrocortisone induced animal model.J Proteome Res,2005,4,(6):2391
[15] Brindle JT, Antti H, Holmes E,et al.Rapid and noninvasive diagnosis of the presence and severity of coronary heart disease using 1HNMR-based metabonomics.Nat Med,2002,8(12):1439
[16] Nicholson JK,Lindon JC,Holmes E. Metabonomics: understanding themetabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biologica NMR spectroscopic data.Xenobiotica,1999,29(11): 1181-1189
[17] Griffin JL,BollardM E. Metabonomics: Its potentialas a tool in toxicology for safety assessment and data integration.CuDrug Metab,2004,5(5): 389-398
[18] Beckwith-Hall BM,Holmes E,Lindon JC,et al. NMR-based metabonomic studies on the biochemical effects of commonly used drug carrier vehicles in the rat.Chem ResToxicol,2002, 15(9): 1136-1141
[19] Lindon JC,Nicholson J K,Holmes E,et al. Metabonomics: Metabolic processes studied by NMR spectroscopy of biofluids.Concept Magnetic Res,2000,12(5): 289-320
[20] Nicholson JK,Wilson ID. High resolution proton NMR spectroscopy of biological fluids.Prog. NMR Spectrosc. 1989,21,444-501
[21] Solanky KS,BaileyN JC,Holmes E,et al. NMR-based metabonomic studies on the biochemical effects of epicatechin in rat.J Agric Chem,2003,51(14): 4139-4145
[22] Lafaye A,Junot C,Ramounet-LeGall B,et al. Metabolite profi-ling in rat urine by liquid chromatography/electrospray on trap mass spectrometry..Rapid Common Mass Spectrom,2003, 17: 2541-2549
[23] Thomp som JA, Markey SP. Quantitative metabolic profiling of urinary organic acids by gaschromatography-mass spectrometry: comparison of isolation methods.Anal Chem, 1975, 47 (8): 1313-1321
[24] Niwa T.Metabolic profiling with gas chromatography-mass spectrometry and its application to clinical medicine. J Chromatogr, 1986, 379: 313-345
[25] Holmes E,Nicholls AW,Lindon JC,et a1.Chemometrie models for toxicity classification based on NMR spectra of biofluids.Chem Res Toxico,2000,13(6);471-478
[26] Robertson DG,Reily MD,Sigler RE,et a1.Metabonomics:evaluation of nuclear magnetic resonance (NMR)and pattern recognition technology for repid in vivo screening of liver and kidney toxicants.Toxicol Sci,2000,57(2):326-337
[27]颜贤忠,赵剑宇,彭双清,廖明阳.代谢组学在后基因组时代的作用.波谱学杂志,2004,21 (2):263-267
[28] Slim RM, Robertson DG, AlbassamM,et al. Effect of dexamethasone on the metabonomics profile associated with phosphodiesterase inhibitor-induced vascular lesions in rats. Toxicol Appl Pharm,2002, 183(2): 108-109
[29]王全军,颜贤忠,吴纯启,等. Z24经口染毒大鼠血浆的代谢组学研究.卫生毒理学杂志,2004, 18(2):74-76
[2]邓培嫒,傅琪.116例药源性肝损害分析.药物流行病学杂志,1998,7(3):144-145.
[3]徐列明.对中草药引起肝损害的认识及防治.现代医药卫生,2005,21(4):379-380.
[4]陈勇,程明,刘春霞.肝脏毒性中药的研究现状与展望.中草药,2004,35(11):1315-1317.
[5]傅肖岩,劳绍贤.重视中药所致肝损害.中药新药与临床药理,2003,14(2):130-133.
[6]胡锡琴.对中草药引起药源性肝损害的分析.陕西中医,2002,23(9):837-838.
[7]卢晓露,张煜,赵肃清.具肝毒性天然药物的研究进展及防治策略.现代食品与药品杂志,2007,17(1):11-15.
[8]中华人民共和国药典委员会.中华人民共和国药典[S].一部.北京:化学工业出版社,2005:111-112.
[9]李全林,朱桂芳.复方苍耳子油剂治疗鼻窦炎98例.陕西中医,2009,30(12):1644.
[10]骆桂秋,顾析玲,宋宪华.苍耳子汤配合西药治疗小儿急性鼻窦炎73例—附单纯西药治疗62例对照.浙江中医杂志,2005,40(7):289-289.
[11]李红,周谋.苍耳子及复方制剂的药理作用和临床研究进展.山西医科大学学报,2004,35(3):313-314.
[12]华刚,孟静.玉屏风散合苍耳子散加减治疗过敏性36例.河南中医,2005,25(3):65-65.
[13]侯自强.连蒲苍耳子散联合药物冲洗治疗慢性鼻窦炎138例疗效观察.中医药导报,2009,15(8):52-54
[14]韩婷.苍耳子的生物活性成分及品质评价(博士论文).上海:第二军医大学,2006.
[15]韩婷,秦路平,郑汉臣.苍耳及其同属药用植物研究进展.解放军药学学报,2003,19(2):122-125.
[16]Lu-ping Qin,Ting Han,Huiliang Li,Qiaoyan Zhang,et al. A new thiazinedione from Xanthium strumarium. Fitoterapia, 2006,77: 245-246.
[17]Ting Han,Huiliang Li,Qiaoyan Zhang,et al. New thiazinediones and other components from Xanthium strumarium L.. Chem Nat Compd, 2006,42(5): 567-570.
[18]张晓琦.苍耳属中倍半萜内酯的研究进展.中草药,2001,32(10):951-953.
[19]梅全喜,毕焕新.现代中药药理手册.北京:中国中医药出版社,1998,41-43.
[20]宋振玉,张凌云,谢明智,等.苍耳子有毒成分及药理作用.药学学报,1962,9(11):678-683.
[21]何芙莉,闫姝,张小波.苍耳子小鼠最大耐受量实验观察.中国中西医结合外科杂志, 1998,4(2):107-108.
[22]MacLeod JK,Moeller PD,Franke FP. Two toxic glycosides from the burrs of Xanthium pungens. J Nat Prod, 1990, 53(3): 451.
[23]王素贤,任丽娟,孙泽人.蒙古苍耳种仁中的有毒成分.中草药,1983,14(12):1.
[24]苏亚,翁维良,胡瑾.临床中药学.郑州:河南科技出版社,1998,214-215.
[25]张学梅,张重华.苍耳子中毒及毒性研究进展.中西医结合学报.2003,1(1):71-73.
[26]郭汉林.急性鲜苍耳子中毒12例报告.临床内科杂志,1989,6(2):29.
[27]周加权.以多器官损害为表现的苍耳子中毒1例报告.中国社区医师,2005,21(5):47.
[28]赵胜乾,吴敏.苍耳子中毒致急性肾功能衰竭及肝损伤1例.实用诊断与治疗杂志,2004,18(6):514.
[29]梁晓燕,朱会友,王建丽.小剂量苍耳子中毒1例.实用中医药杂志,1998,14(10):45-46.
[30]丁雅芳.简述中药毒副反应.湖北中医杂志,1985,4:43.
[31]纪绍先,张国栋.急性苍耳子中毒12例报告.吉林医科大学学报,1962,1(1):75-78.
[32]杜青云,潘东林.73例急性中药中毒分析.药学实践杂志,1995,13(5):277-279.
[33]王希海,黄光照.药用植物引起中毒性肝病时的病理变化.中西医结合肝病杂志,1996, 6(4):48-50.
[34]朱天忠.试论中草药的肝毒作用.中国法医学杂志,1986,1(1):56.
[35]王文英.部分中药对肾脏毒副作用及防治.时珍国医国药,2000,11(4):347-348.
[36]赵宝玉,王建华,史克诚.动物毒理学.北京:中国农业出版社,2001,349-351.
[37]钱培忠.有毒中药大辞典.天津:天津科技出版公司,1995,258-260.
[38]姜萧,隋进江.浅谈中药苍耳子毒性及中毒的抢救方法.黑龙江中医药,2000,20(4):63.
[39]李涓,高天,谢子清,唐熠.苍耳子不同提取物的毒性比较实验.时珍国医国药,2005,16(6)484-488.
[40]Nishimura H,Nonaka GI,Nishioka I. Seven quinic acid gallates from Quercus stenophylla. Phytochemistry,1984,23 (11):2621-2623.
[41]Morishita H, et a1.Chromatographic separation and identification of naturally occurring choirogenic acids by 1H NMR spectroscopy and mass spectrometry. J Chromatogr 1984,315:253.
[42]郑晓珂,李钦,冯卫生.冬凌草中酚酸类化学成分研究.中国药学杂志,2004,39(5):335-336.
[43]Ma YT, Huang MC, Hsu FL, et al. Thiazinedion from Xanthium strumarium. Phytochemistry, 1998, 48(6): 1083-1085.
[44]Mahmoud,Ahmed A, Al-Shihry, Shar S, et al. A new heterocyclic glucoside from the fruits of Xanthium pungens.Natural Product Research,2005,19(6): 585-589.
[45]Sonia P,Cosimo P,Nunziatina DT,et al. Sesquiterpene and diterpene glycosides from Xanthium spinosum.Phytochemistry,1996,41(5):1357-60.
[46]翁维良.临床中药不良反应.中药新药与临床药理,1996,7(2):4.
[47]雷树勇,黄承乐,陆爱权.11种生化项目在肝脏疾病中的变化分析.2002,14(1):59-61.
[48]刘娓娓,顾文君,沈锡中.血清生化指标对慢性乙型肝炎肝纤维化及炎症程度的诊断价值.中国临床医学,2005,12(1):65-66.
[49]周加权.以多器官损害为表现的苍耳子中毒1例报告.中国社区医师,2005,21(5):47.
[50]叶秀珍,朱德厚,沈鼎武.体外连续培养的成人肝细胞的超显微结构.实验生物学报, 1980,13:3612-3641.
[51]郭竹英,矫强,徐芒华,王世婷.脂多糖对人正常肝细胞株L02损伤的实验研究.医学分子生物学杂志,2008,5(4):323-327.
[52]陈莹,季莉莉,马鸿雁,刘天瑜,王峥涛.吡咯里西啶生物碱Jacoline对人正常肝L-02细胞内谷胱甘肽含量及还原酶活性的影响.上海中医药大学学报.2008,22(2):35-39.
[53]孟雪莲,曾晓非,孙祖越,裴晓丹.丹参致大鼠肝细胞脂质过氧化损伤的实验研究.辽宁大学学报,2005,32(1):31-34.
[54]周成江,周立社,和彦苓,杨艳芳.蒙药童格勒格四种粗提物对大鼠正常肝细胞BRL株增殖的影响.包头医学院学报,2008,24(4):348-349.
[55]陈国勇,曲青山,孙建军,钱建民.转染血红素氧化酶21基因大鼠肝细胞系BRL体外抗凋亡作用.郑州大学学报(医学版),2006,41(4):712-714.
[56]马巧玉,王宇明,郝飞.LDH检测BALB/c小鼠CTL活性的初步研究.第三军医大学学报, 2003,25(4):298-300.
[57]王甫厚,陈绍先,郭彩云,等.LDH释放法测NK细胞毒的方法学研究.中国免疫学杂志, 1990,6(2):115.
[58]张蕾,李芳秋,张士新,黄海嵘,李德闵.LDH释放法检测肿瘤患者PBMC和CIK细胞的细胞毒活性.现代检验医学杂志,2008,23(6):27-29.
[59]Cole RJ,Stuart BP,Lansden JA,Cox RH.Isolation and redefinition of the toxic agent fromcocklebur (Xanthium strumarium). J Agric Food Chem,1980,28: 1330-1332.
[60]Georgiou M,Sianidon L,Hatziz T,Papadatos J,Konselini SA. Hepatotoxicity due to Atractylis gummifera L.Clin Toxicol 1988,26:487-493.
[61]Larrey D.Hepatotoxicity of herbal remedies.J Hepatol 1997,26:47-54.
[62]Mokhobo KP.Herb use and necro-degenerative hepatitis.S Afr Med J 1976,50:1096-9.
[63]Luciani S,Carpenedo F,Tarjan EM. Effects of atractyloside and carboxyatracty-loside in the whole animal. In Atractyloside: Chemistry, Biochemistry and Toxicology1978,109.
[64]Obatomi DK, Bach PH.Biochemistry and Toxicology of the Diterpenoid Glycoside Atractyloside.Food and Chemical Toxicology 1998,36:335-346.
[65]Sinclair AJ, et al. NMR-based metabolomic analysis of cerebrospinal fluid and serum in neurological diseases - a diagnostic tool.NMR Biomed, 2010,23(2):123-132.
[66]Lee JE, et al.Evidence of vintage effects on grape wines using 1H NMR-based metabolomic study.Anal Chim Acta,2009.648(1):71-76.
[67]Lee IJ, et al. NMR metabolomic analysis of caco-2 cell differentiation. J Proteome Res, 2009,8(8):4104-4108.
[68]Ko BK, et al. Metabolomic insight into soy sauce through 1H NMR spectroscopy. J Agric Food Chem,2009,57(15):6862-6870.
[69]Hirakawa K, et al. Experimental estimation of postmortem interval using multivariate analysis of proton NMR metabolomic data. Leg Med (Tokyo),2009,11(1):282-285.
[70]Nicholas PC, et al. 1H NMR-based metabolomic analysis of liver,serum,and brain following ethanol administration in rats.Chem Res Toxicol,2008,21(2):408-420.
[71]Kruger NJ, Troncoso-Ponce MA, Ratcliffe RG. 1H NMR metabolite fingerprinting and metabolomic analysis of perchloric acid extracts from plant tissues. Nat Protoc,2008,3(6):1001-1012.
[72]Poulding S, et al. Removal of t(1) noise from metabolomic 2D(1)H-(13)C HSQC NMR spectra by Correlated Trace Denoising.J Magn Reson, 2007.189(2): 190-199.
[73]Gu H, et al. 1H NMR study of the effects of sample contamination in the metabolomic analysis of mouse urine.J Pharm Biomed Anal,2007,45(1):134-140.
[74]Cho IH, Kim YS, Choi HK. Metabolomic discrimination of different grades of pine mushroom (Tricholoma matsutake Sing.) using 1H NMR spectrometry and multivariate dataanalysis.J Pharm Biomed Anal,2007,43(3):900-904.
[75]Zhou J, et al.1H NMR-based metabonomic and pattern recognition analysis for detection of oral squamous cell carcinoma.Clin Chim Acta,2009,401(1):8-13.
[76]Xiao C, et al. Revealing the metabonomic variation of rosemary extracts using 1H NMR spectroscopy and multivariate data analysis.J Agric Food Chem,2008,56(21): 142-153.
[77]Craig A, et al.Scaling and normalization effects in NMR spectroscopic metabonomic data sets.Anal Chem,2006,78(7):2262-2267.
[78]Webb-Robertson BJ, et al. A study of spectral integration and normalization in NMR-based metabonomic analyses.J Pharm Biomed Anal,2005,39(3-4):830-836.
[79]Bundy JG, et al. A NMR-based metabonomic approach to the investigation of coelomic fluid biochemistry in earthworms under toxic stress.FEBS Lett,2001,500(1-2):31-35.
[80]Chan ECY, et al. Metabolic Profiling of Human Colorectal Cancer Using High–Resolution Magic Angle Spinning Nuclear Magnetic Resonance (HR-MAS NMR) Spectroscopy and Gas Chromatography Mass Spectrometry (GC/MS).J Proteome Res,2009,8(1):352-361.
[81]Yamamoto T, Horii I, Yoshida T.Integrated NMR-based metabonomic investigation of early metabolic effects of ethylene glycol monomethyl ether (EGME) on male reproductive organs in rats.J Toxicol Sci,2007,32(5):515-528.
[82]Psihogios NG, et al.Evaluation of tubulointerstitial lesions severity in patients with glomeru-onephritides:an NMR-based metabonomic study. J Proteome Res,2007, 6(9):3760-3770.
[83]Bertram HC,et al. Effect of magnetic field strength on NMR-based metabonomic human urine data.Comparative study of 250,400,500,and 800 MHz. Anal Chem, 2007,79(18): 7110-7115.
[84]Bertram HC,et al. NMR-based metabonomic studies reveal changes in the biochemical profile of plasma and urine from pigs fed high-fibre rye bread. Br J Nutr, 2006. 95(5): 955-962.
[85]Zha W, et al.Metabonomic characterization of early atherosclerosis in hamsters with induced cholesterol. Biomarkers,2009.14(6):372-380.
[86]Yin P, et al.A metabonomic study of hepatitis B-induced liver cirrhosis and hepatocellular carcinoma by using RP-LC and HILIC coupled with mass spectrometry.MolBiosyst,2009,5(8):868-876.
[87]Sieber M, et al. Metabonomic Study of Ochratoxin A Toxicity in Rats after Repeated Administration:Phenotypic Anchoring Enhances the Ability for Biomarker Discovery.Chem Res Toxicol,2009,22(7):1221-1231.
[88]Yan S, et al. Metabonomic characterization of aging and investigation on the anti-aging effect of total flavones of Epimedium.Mol Biosyst, 2009,5(10):1204-1213.
[89]Huang X, et al.A metabonomic characterization of CCl4-induced acute liver failure using partial least square regression based on the GC/MS metabolic profiles of plasma in mice.J Chromatogr B Analyt Technol Biomed Life Sci,2008,870(2):178-185.
[90]Fardet A, et al.Whole-grain and refined wheat flours show distinct metabolic profiles in rats as assessed by a 1H NMR-based metabonomic approach. J Nutr,2007.137(4):923-929.
[91]Keun HC. Metabonomic modeling of drug toxicity. Pharmacology Therapeutics, 2006, 109(1-2):92-106.
[92]Beckonert O, et al. NMR-based metabonomic toxicity classification: hierarchical cluster analysis and k-nearest-neighbour approaches. Anal. Chim. Acta,2003.490(1-2): 3-15.
[93]Nicholls AW, et al. Metabonomic investigations into hydrazine toxicity in the rat. Chem Res Toxicol.,2001,14(8):975-987.
[94]Bollard ME, et al.Comparative metabonomics of differential hydrazine toxicity in the rat and mouse.Toxicol.Appl Pharmacol,2005,204(2):135-151.
[95]Mortishire-Smith RJ, et al.Use of metabonomics to identify impaired fatty acid metabolism as the mechanism of a drug-induced toxicity.Chem Res Toxicol,2004,17(2):165-173.
[96]Ebbels T, et al.Toxicity classification from metabonomic data using a density superposition approach:'CLOUDS'.Anal Chim Acta,2002,490(1-2):109-122.
[97]Wu H, et al. NMR spectroscopic-based metabonomic investigation on the acute biochemical effects induced by Ce(NO3)3 in rats. J Inorg Biochem,2005,99(11):2151-2160.
[98]Zhang XY, et al. NMR-based metabonomic study on the subacute toxicity of aristolochic acid in rats.Food and Chemical Toxicology,2006,44(7):1006-1014.
[99]李中港,程芳,盛卸晃,刘兆平.代谢组学在药物安全性评价中的应用进展.毒理学杂志. 2005,24(5):415-418.
[100]王全军,颜贤忠,吴纯启.Z24经口染毒大鼠尿液的核磁共振谱代谢组学研究.中国药理学与毒理学杂志,2004,18(6):465-470.
[101]王全军,颜贤忠,吴纯启.Z24经口染毒大鼠血浆的代谢组学研究.卫生毒理学杂志,2004,18(2):73-75.
[102]李伶,颜贤忠.代谢组学在毒理学中的应用进展.军事医学科学院院刊,2007,31(2): 183-186.
[103]何君,周宏灏.代谢组学及其在药理学中的进展.中国药理学通报,2006,22(11):1304-1309.
[104]Nicholson JK,Conelly J,Lindon JC,et al. Metabonomics:A Platform for studying drug toxicity and gene function.Nat Rev Drug Discov,2002,1(2):153-161.
[105]Holmes E, Nicholls AW, Lindon JC,et al. A development of a model for classification of toxin-induced lesions using lH NMR spectroscopy of urine combined with pattern recognition.NMRBiomed,l998,11(4-5):235-244.
[106]Holmes E,Nicholson JK, NiCholls AW,et al.The identification of novel Biomarkers of renal toxicity using automatic data reduction techniques and PCA of Proton NMR spectra of urine.Chemomet.lntel Lab systems,1998,44:245-255.
[107]Holmes E,Nicholls AW,Lindon JC,et al.Chemometric models for toxicity classification based on NMR spectra of biofluids.Cheml Res Toxicol,2000,13(6):471-478.
[108]Nicholson JK,lindon J,Holmes E.Metabonomics:understanding the Metabolic responses of living systems to pathophysiological stimuli via muItivariate statisticaI analysis of biological NMR spectroscopic data Xenobiotica,1999,29(11):1181-1189.
[1] Nicholson JK,Wilson ID. Opinion:understanding global systems biology:metabonomics and the continuum of metabolism.Nat Rev Drug Discov,2003,2(8):668-676
[2] Fiehn O. Combining genomics, metabolome analysis, and biochemical modelling to understand metabolic networks. Comp Funct Genom, 2001, 2: 155-168
[3] Fiehn O. Metabolomics-the link between genotypes and phenotypes.Plant Mol Biol, 2002, 48: 155-171
[4] Nicholson JK, Connelly J, Lindon JC,et al. Metabonomics: a plat-form for studying drug toxicity and gene function.Nat Rev Drug Discov,2002,1(2):153
[5] Lindon JC,Nicholson JK,Holmes E. Metabonomics: Metabolic processes studied by NMR spectroscopy of biofluids. Concepts Magn Reson,2000,12,5:289-320
[6] Lindon JC, Holmes E, Nicholson JK. Recent development and Applications of NMR-Based Metabonomics. Chinese Journal of Magnetic Resonance, 2006,23(1):101-127
[7] Plumb RS,Stumpf CL,Granger JH,et al. Use of liquid chro-matography/time-of-flight mass spectrometry and multivariate statistical analysis shows promise for the detection of drug metabolites in biological fluids.Papid Communic in Mass Spectrome,2003,17(23): 2632-2638
[8] Plumb RS,Stumpf CL,Gorenstein MV, et al. Metabonomics:the use of electrospray mass spectrometry coupled to reversed phase liquid chromatography shows potential for the screening on rat urine in drug development.Rapid Communic in MasSpectrome, 2002, 16(20): 1991-1996
[9] Beaudry F,Le Blanc JC Y,Coutu M,et al. Metabolite profiling study of propranolol in ratusing LC/MS/MS analysis.Biomed Chromatogra,1999,13(5): 363-369
[10] Hai PT, Kaskavelis L, Daykin CA, Janssen HJ. Method development in high-performance liquid chromatography for high-throughput profiling and metabonomic studies of biofluid samples. Chromatogr B2003, 789(2): 283-301
[11] Plumb RS, Stumpf CL, Gorenstein MV, Castro-Perez JM, Dear GJ, Anthony M, Sweatman BC, Connor SC, Haselden JN. Metabonomics: the use of electrospray mass spectrometry coupled to reversed-phase liquid chromatography shows potential for the screening of rat urine in drug development.Rapi Commun Mass Spec, 2002, 16: 1991-1996
[12] Bugrim A,Nikolskaya T,Nikolsky Y. Early prediction of drug metabolism and toxicity: systems biology approach and modeling.Drug DiscovT 2004,9(3): 127-135
[13] Taylor J, King RD, AltmannT, et al. Application of metabolomics to plant genotype discrimination using statistics and machine learning.Bioinformatics,2002,18(2): 241
[14] Chen MJ, Zhao LP, Jia W.Metabonomic study on the biochemical profiles of a hydrocortisone induced animal model.J Proteome Res,2005,4,(6):2391
[15] Brindle JT, Antti H, Holmes E,et al.Rapid and noninvasive diagnosis of the presence and severity of coronary heart disease using 1HNMR-based metabonomics.Nat Med,2002,8(12):1439
[16] Nicholson JK,Lindon JC,Holmes E. Metabonomics: understanding themetabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biologica NMR spectroscopic data.Xenobiotica,1999,29(11): 1181-1189
[17] Griffin JL,BollardM E. Metabonomics: Its potentialas a tool in toxicology for safety assessment and data integration.CuDrug Metab,2004,5(5): 389-398
[18] Beckwith-Hall BM,Holmes E,Lindon JC,et al. NMR-based metabonomic studies on the biochemical effects of commonly used drug carrier vehicles in the rat.Chem ResToxicol,2002, 15(9): 1136-1141
[19] Lindon JC,Nicholson J K,Holmes E,et al. Metabonomics: Metabolic processes studied by NMR spectroscopy of biofluids.Concept Magnetic Res,2000,12(5): 289-320
[20] Nicholson JK,Wilson ID. High resolution proton NMR spectroscopy of biological fluids.Prog. NMR Spectrosc. 1989,21,444-501
[21] Solanky KS,BaileyN JC,Holmes E,et al. NMR-based metabonomic studies on the biochemical effects of epicatechin in rat.J Agric Chem,2003,51(14): 4139-4145
[22] Lafaye A,Junot C,Ramounet-LeGall B,et al. Metabolite profi-ling in rat urine by liquid chromatography/electrospray on trap mass spectrometry..Rapid Common Mass Spectrom,2003, 17: 2541-2549
[23] Thomp som JA, Markey SP. Quantitative metabolic profiling of urinary organic acids by gaschromatography-mass spectrometry: comparison of isolation methods.Anal Chem, 1975, 47 (8): 1313-1321
[24] Niwa T.Metabolic profiling with gas chromatography-mass spectrometry and its application to clinical medicine. J Chromatogr, 1986, 379: 313-345
[25] Holmes E,Nicholls AW,Lindon JC,et a1.Chemometrie models for toxicity classification based on NMR spectra of biofluids.Chem Res Toxico,2000,13(6);471-478
[26] Robertson DG,Reily MD,Sigler RE,et a1.Metabonomics:evaluation of nuclear magnetic resonance (NMR)and pattern recognition technology for repid in vivo screening of liver and kidney toxicants.Toxicol Sci,2000,57(2):326-337
[27]颜贤忠,赵剑宇,彭双清,廖明阳.代谢组学在后基因组时代的作用.波谱学杂志,2004,21 (2):263-267
[28] Slim RM, Robertson DG, AlbassamM,et al. Effect of dexamethasone on the metabonomics profile associated with phosphodiesterase inhibitor-induced vascular lesions in rats. Toxicol Appl Pharm,2002, 183(2): 108-109
[29]王全军,颜贤忠,吴纯启,等. Z24经口染毒大鼠血浆的代谢组学研究.卫生毒理学杂志,2004, 18(2):74-76