雷公藤致大鼠肾毒性血清代谢组学分析

目的:联合超高效液相色谱与复合型三重四级杆/线性离子阱串联质谱联用技术(UFLC-MS/MS)及统计学分析方法研究雷公藤对大鼠血清中内源性小分子化合物的影响和肾毒性作用机制,寻找与肾毒性相关的生物标记物和代谢通路。方法:利用试剂盒法测定给药14 d后大鼠血清生化指标,观察肾组织切片。血清样本采用乙腈沉淀蛋白法,UFLC-MS/MS分析测定,正离子一级全扫描模式,经主成分分析法和主成分判别分析法处理数据。结果:与空白组大鼠相比,雷公藤乙醇提取物组大鼠血清生化指标(血清肌酐、尿素氮)显著升高;组织病理学切片明显病变,表明造模成功。血清中二甲基甘氨酸、赖氨酸、苯丙氨酸、溶血卵磷脂、磷脂酰胆碱、磷脂酰丝氨酸和甘油二酯等9种代谢物有显著性差异,鉴定为雷公藤致肾毒性的生物标记物。结论:雷公藤对肾脏的毒性作用机制可能与氨基酸代谢、磷脂代谢等有关,为雷公藤类药物临床毒性的早期预防提供参考。

Metabonomic Analysis on Serum in Nephrotoxic Rats Induced by Radix et Rhizoma Tripterygii

Objective: An ultrafast liquid chromatography-hybrid triple quadrupole/linear ion trap mass spectrometry (UFLC-MS/MS) combined with statistical analysis method was used to investigate the changes of endogenous small molecule compounds and nephrotoxic mechanism in the rat serum induced by Radix et Rhizoma Tripterygii and to explore biomarkers associated with renal toxicity as well as metabolism pathways. Method: After intragastric administration for 14 days,serum biochemical indexes was detected by kit method and nephridial tissue section was observed.The serum samples were added acetonitrile for protein precipitation and analyzed by UFLC-MS/MS with full scan mode under positive ion scan mode,then principle component analysis (PCA) and principle component analysis discriminant analysis (PCA-DA) were used for data processing. Result: Compared with the blank group,serum biochemical indexescreatinine (Cr) and blood urea nitrogen (BUN)] of Radix et Rhizoma Tripterygii ethanol extract group increased significantly and histopathological section damaged obviously which showed that the animal module prepared well.Nine significant different metabolites, such as dimethylglycine,lysine,phenylalanine,lysolecithin,phosphatidyl choline,phosphatidylserine and diglyceride, were identified as biomarkers of nephrotoxicity induced by Radix et Rhizoma Tripterygii. Conclusion: Nephrotoxic mechanism induced by Radix et Rhizoma Tripterygii may be related to the metabolism of amino acid and phospholipids,and it provides reference relating to the early clinical toxicity prevention of Radix et Rhizoma Tripterygii.