基于液质联用技术的金铃子散亚急性毒性代谢组学(1)
[摘要] 为进一步了解金铃子散毒性影响的代谢特征，探索金铃子散在治疗过程中对未知生物通路的作用。该实验通过以金铃子散3组剂量的水煎剂对大鼠肝肾的影响，来研究金铃子散对大鼠体内代谢组改变的特点和规律。首先对给药2周和给药后1周大鼠尿液和血清样本采用质谱分析方法，通过XCMS在线分析大鼠正负模式离子检测谱，对代谢谱分析的代谢物进行鉴定，结果得到44个差异性物质。最后以MetPA对液质实验中筛选出的代谢物组进行代谢通路富集和分析，结果得到：类固醇激素合成代谢途径，戊糖和葡萄糖醛酸转换，赖氨酸代谢，谷胱甘肽代谢和抗坏血酸代谢5条相关的代谢通路。 由代谢网络图可以看出，与毒性相关的通路主要和生物体赖氨酸代谢、葡萄糖醛酸转化及激素代谢有关，尤其是赖氨酸和谷胱甘肽的代谢失衡，造成机体能量代谢或氧化应激调节的紊乱，产生大鼠体内的损伤。从金铃子散低、中、高3组剂量的亚急性毒性实验结果观察发现，金铃子散19.7 g·kg-1和 39.4 g·kg-1剂量组造成了明显的毒性作用，说明金铃子散在生物体内可产生毒性作用，且毒性随着给药剂量的增加而增加，造成不可逆的机体损伤。
, http://www.100md.com
[关键词] 金铃子散； 毒性； 代谢组学； 液质联用
[Abstract] To further understand the metabolic characteristics of Jinlingzi powder toxicity effect in rats and explore the effect of Jinlingzi powder on unknown biological pathways in the treatment process. In this experiment， the effect of three doses of Jinlingzi powder decoction on rat liver and kidney was investigated to explore the characteristics and rules of Jinlingzi powder on in vivo metabonomic changes in rats. First， urine and serum samples of the rats were used for LC-MS analysis. Under the XCMS online analysis， 44 differential substances were found in the identification of metabolites. Finally， Metpa was used for metabolic pathways enrichment and analysis， and five related metabolic pathways were obtained： steroid hormone biosynthesis， tryptophan metabolism， pentose and glucuronate interconversions， ascorbate and aldarate metabolism， as well as glutathione metabolism. Metabolic network diagram showed that the toxicity-related pathways were mainly associated with lysine metabolism in living organisms， glucuronic acid conversion， and hormone metabolism， especially the metabolism imbalance of lysine and glutathione would result in the disorder of energy metabolism or oxidative stress regulation， and thus inducing the damage in rats. Subacute toxicity test results for three doses groups (low， middle and high doses) showed that， Jinlingzi powder with doses of 19.7 g·kg-1 and 39.4 g·kg-1 caused obvious toxic effect， indicating Jinlingzi powder could produce toxic effect in vivo in a dose-dependent manner， and cause irreversible damage to the body.
, 百拇医药
[Key words] Jinlingzi powder； toxicity； metabonomics； LC-MS
代謝组学主要研究机体小分子代谢物与疾病的关系以及揭示机体内生物化学变化的规律。将这样的方法应用在中药毒性的研究中可以较容易的发现毒性作用的物质基础和作用机制，对追踪毒性作用的相关代谢途径，寻找代表性的生物标志物有重要作用[1-2]。
疏肝泄热名方金铃子散由川楝子和延胡索组成，而两药均有一定的肝肾毒性，尤其是川楝子早在《名医别录》就有记载“楝实有小毒。根微寒。治蛔虫， 利大肠。生荆山” [3]。 但作为中药复方金铃子散毒性的探索并没有报道，为进一步明确金铃子散对机体肝肾代谢机制的影响和产生的毒性作用，本实验从代谢组学的视角对金铃子散作用于肝肾毒性的影响以及毒性标志物的确定进行了初步研究。
1 材料与方法, 百拇医药(沈淑洁 水素芳 肖炳坤 杨建云 黄荣清)
, http://www.100md.com
[关键词] 金铃子散； 毒性； 代谢组学； 液质联用
[Abstract] To further understand the metabolic characteristics of Jinlingzi powder toxicity effect in rats and explore the effect of Jinlingzi powder on unknown biological pathways in the treatment process. In this experiment， the effect of three doses of Jinlingzi powder decoction on rat liver and kidney was investigated to explore the characteristics and rules of Jinlingzi powder on in vivo metabonomic changes in rats. First， urine and serum samples of the rats were used for LC-MS analysis. Under the XCMS online analysis， 44 differential substances were found in the identification of metabolites. Finally， Metpa was used for metabolic pathways enrichment and analysis， and five related metabolic pathways were obtained： steroid hormone biosynthesis， tryptophan metabolism， pentose and glucuronate interconversions， ascorbate and aldarate metabolism， as well as glutathione metabolism. Metabolic network diagram showed that the toxicity-related pathways were mainly associated with lysine metabolism in living organisms， glucuronic acid conversion， and hormone metabolism， especially the metabolism imbalance of lysine and glutathione would result in the disorder of energy metabolism or oxidative stress regulation， and thus inducing the damage in rats. Subacute toxicity test results for three doses groups (low， middle and high doses) showed that， Jinlingzi powder with doses of 19.7 g·kg-1 and 39.4 g·kg-1 caused obvious toxic effect， indicating Jinlingzi powder could produce toxic effect in vivo in a dose-dependent manner， and cause irreversible damage to the body.
, 百拇医药
[Key words] Jinlingzi powder； toxicity； metabonomics； LC-MS
代謝组学主要研究机体小分子代谢物与疾病的关系以及揭示机体内生物化学变化的规律。将这样的方法应用在中药毒性的研究中可以较容易的发现毒性作用的物质基础和作用机制，对追踪毒性作用的相关代谢途径，寻找代表性的生物标志物有重要作用[1-2]。
疏肝泄热名方金铃子散由川楝子和延胡索组成，而两药均有一定的肝肾毒性，尤其是川楝子早在《名医别录》就有记载“楝实有小毒。根微寒。治蛔虫， 利大肠。生荆山” [3]。 但作为中药复方金铃子散毒性的探索并没有报道，为进一步明确金铃子散对机体肝肾代谢机制的影响和产生的毒性作用，本实验从代谢组学的视角对金铃子散作用于肝肾毒性的影响以及毒性标志物的确定进行了初步研究。
1 材料与方法, 百拇医药(沈淑洁 水素芳 肖炳坤 杨建云 黄荣清)