基于“异类相制”的马钱子复方配伍减毒的代谢组学研究

Metabolomics of Strychnos about Formulation Detoxicity Effect Based on "Heterogeneous Restriction" Theory

目的:马钱子是通络止痛的要药,但其不良反应临床上常有发生,基于中医"异类相制"理论探讨马钱子复方配伍减毒,以期为临床合理用药提供参考。方法:采用血清生化检测、肾组织病理HE染色技术,观察血清生化指标CREA和UREA以及肾脏组织病理变化,并利用UPLC-TOF-MS分析方法获得马钱子组、健脾通络复方组和对照组大鼠血清代谢轮廓,运用正交偏最小二乘法-判别分析(OPLS-DA)和变量重要性投影(VIP)值筛选差异性代谢产物。结果:马钱子组中血清CREA和UREAL上调,HE染色显示马钱子组大鼠肾组织病变严重,健脾通络复方组能逆转此趋势,改善病变。UPLC-TOF-MS分析各组大鼠血清代谢轮廓获得这些肾毒性生物标志物主要有喹啉酸、L-脯氨酸、肌酸酐、吲哚乙酸、胍基丁二酸、同型半胱氨酸、L-精氨酸、柠檬酸、牛磺酸,代谢通路富集分析关联到牛磺酸和次牛磺酸代谢、精氨酸和脯氨酸代谢、精氨酸生物合成、三羧酸循环、色氨酸代谢等主要代谢通路。结论:该研究从代谢的角度阐明马钱子肾毒性机制可能与氨基酸代谢异常、能量代谢紊乱、氧化应激有关,而配伍后的健脾通络方对肾脏具有明显减毒作用,科学揭示了中医配伍减毒的科学内涵。

Objective: To study the detoxicity effect of Strychnos in TCM compound formula based on the "heterogeneous restriction" of TCM theory, so as to provide reference for improving the rationality of its clinical application. Methods: Serum biochemistry detection and renal histopathological HE staining were used to observe the changes of serum biochemical indexes of CREA and UREA, as well as the kidney tissue pathological. The serum metabolic profile of rats was performed by using UPLC-TOF-MS to analyze the metabolic profiles of Strychnos, Jianpi Tongtuo Compound Formula group and the control group. The OPLS-DA score plots and VIP value were used to screen out the differential metabolites. Results: The levels of CREA and UREA were significantly up-regulated in the Strychnos group, and the renal injury of rats in the Strychnos group were severe by HE staining. This trend could be reversed and the lesion could be improved in the Jianpi Tongluo Compound Formula group. UPLC-TOF-MS was used to analyze the serum metabolic profile of rats in each group. The main biomarkers of nephrotoxicity, including quinolinic acid, L-proline, creatinine, indoleacetic acid, guanidine succinic acid, homocysteine, L-arginine, citric acid and taurine, were confirmed by multivariate statistical analysis. The metabolic pathways enrichment analysis linked to main metabolic pathways included Taurine and hypotaurine metabolism, Arginine and proline metabolism, Arginine biosynthesis, TCA cycle and Tryptophan metabolism. Conclusion: From the perspective of metabolism, this study elucidated that the renal toxicity mechanism of Strychnos might be related to abnormal amino acid metabolism, energy metabolism disorder, and oxidative stress. Furthermore, Jianpi Tongluo Compound Formula has obvious effect on reducing kidney toxcity with compatibility, it also provides the scientific formulation detoxity connotation of Strychnos.