GC/TOF-MS代谢组学技术研究雷公藤甲素在大鼠体内的急性毒性

采用GC/TOF-MS代谢组学技术研究雷公藤甲素对大鼠尿液中内源性小分子化合物的影响和毒性作用机制，寻找与毒性相关标志物，为雷公藤类药物临床毒性的早期预防和诊断提供依据。SD大鼠随机分为4组，分别为雷公藤甲素高(2.4 mg/kg)、中(1.2 mg/kg)、低(0.6 mg/kg)剂量组及空白对照组。单次灌胃给药后，收集第0、1、3和7天的尿液，GC/TOF-MS进行分析，对尿液中内源性小分子化合物进行鉴定并获取半定量数据，经过主成分分析发现:不同剂量给药和不同时间取样尿液中内源性小分子代谢谱有明显差异，可被清晰区分并呈明显的动态变化轨迹。代谢组学数据分析结果与常规血液生化和组织病理学检查结果基本吻合。研究结果发现，给药引起尿液中的苹果酸、枸橼酸、牛磺酸、谷氨酸、苏氨酸及软脂酸等发生明显改变，提示与雷公藤甲素毒性相关，为潜在的毒性标志物。本研究结果表明，通过测定尿液中小分子代谢物可反映雷公藤甲素对大鼠的毒性，代谢组学技术可以作为药物毒性评价方法。代谢通路分析结果提示，雷公藤甲素对肝脏的毒性机制可能与大鼠肝脏线粒体受损、影响三羧酸循环、氨基酸代谢及脂质代谢有关。

Gas chromatography time-of-flight mass spectrometry based metabolomic approach to evaluate acute toxicity of triptolide in rats

To evaluate toxicity in rats after orally giving triptolide by using the gas chromatography time-of-flight mass spectrometry(GC/TOF-MS)based metabolomics approach. Potential biomarkers identified here paved a way to new implications of clinical early prevention and diagnosis of safety. In this study, SD rats were randomly divided into four groups, that included high dose(2. 4 mg/kg), medium dose(1. 2 mg/kg), low dose(0. 6 mg/kg)and control groups, respectively. Following by single orally dosing, urine, serum and tissue sample were collected, respectively on day 0, day 1, day 3 and day 7 for a subsequent metabolomic bioanalysis. Multivariate data analysis was then used to process the data which indicated excellent separation between different doses and treatment days. The metabolic pattern after treatment with triptolide at high and medium dose visually depicted the occurrence, development and recovery process of toxicity that vividly demonstrated dose and time-dependent toxicity following different dose of triptolide in rats. Metabolomic results were consistent with those obtained from the routine biochemical assays and histopathological characterization. The results indicated that malate, citrate, taurine, glutamate, threonine and stearic acid, etc. could serve as potential toxicity biomarkers. Our study indicated that a metabolomic method established in this study provided a promising and effective approach to evaluate the mechanism of toxicity and the mechanism of hepatotoxicity was related with the impairment mitochondria, tricarboxylic acid cycle, amino acids and free fatty acids metabolism.