基于尿液代谢组学研究肝药酶抑制剂对何首乌致肝损伤的影响-以CYP3A4为例

Effects of CYP450 Enzyme Inhibitors on Liver Injury Induced by Heshouwu(Polygoni Multiflori Radix) Based on Urine Metabonomics: A Case Study of CYP3A4

目的探讨CYP3A4亚型的特异性抑制剂酮康唑对何首乌致肝损伤的影响。方法将21只大鼠随机分为3组,每组7只,即正常对照组,何首乌组以及何首乌+酮康唑组。连续给药28 d。4周后收集大鼠尿液采用UPLC-Q-TOF-MS进行代谢组学研究,同时辅以血清生化指标及组织病理分析。结果血清生化指标显示与正常组相比何首乌组总胆酸和直接胆红素均显著升高,而酮康唑组总胆酸显著降低。肝脏病理结果显示何首乌组较正常组肝脏有轻到中度病理性改变,而何首乌+酮康唑组较何首乌组有所缓解。LC-MS代谢组学技术筛选到13个与何首乌致肝毒性相关的潜在生物标志物以及抑制CYP3A4后显著回调的4种生物标志物;代谢通路分析得到与何首乌肝毒性显著相关的代谢通路分别为苯丙氨酸、酪氨酸和色氨酸的生物合成,苯丙氨酸代谢和甘油磷脂代谢,而抑制CYP3A4后影响到的通路为甘油磷脂代谢通路。结论 CYP3A4被抑制后甘油磷脂代谢的回调可能是何首乌肝毒性缓解的原因。

Objective To investigate the effect of ketoconazole, namely a specific inhibitor of CYP3 A4, on liver injury induced by Heshouwu(Polygoni Multiflori Radix). Methods Twenty-one rats were randomly divided into 3 groups, 7 cases in each group, namely normal control group, Heshouwu(Polygoni Multiflori Radix) group and Heshouwu(Polygoni Multiflori Radix)+ketoconazole group. After 4 weeks, the urine samples were collected and analyzed by UPLC-Q-TOF-MS for metabonomics. Serum biochemical indexes and histopathological were determined. Results The serum biochemical indexes showed that the total cholic acid and direct bilirubin in the Heshouwu(Polygoni Multiflori Radix) group were significantly higher than those in the normal group, while the total cholic acid in the ketoconazole group was significantly lower. The results of liver pathology showed that the liver of the Heshouwu(Polygoni Multiflori Radix) group had mild to moderate pathological changes compared with the normal group, while the Heshouwu(Polygoni Multiflori Radix)+ketoconazole group was better than the Heshouwu(Polygoni Multiflori Radix) group. Thirteen potential biomarkers related to hepatotoxicity induced by Heshouwu(Polygoni Multiflori Radix) and 4 biomarkers with significant callback after inhibition of CYP3 A4 were screened by metabonomics technology. The metabolic pathway analysis showed that the metabolic pathways significantly related to liver toxicity of Heshouwu(Polygoni Multiflori Radix) were phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and glycerophosphatidylcholine metabolism. But the pathway was glycerol phospholipid metabolism pathway when CYP3 A4 was inhibited. Conclusion The improvement of glycerophosphatidylcholine metabolism after CYP3 A4 inhibition may be the reason of alleviating hepatotoxicity of Heshouwu(Polygoni Multiflori Radix).