基于血浆代谢组学研究制何首乌改善小鼠肾虚的作用机制

Effect and mechanism investigation on improving kidney deficient in mice of Polygoni Multiflori Radix Praeparata based on plasma metabolomics

运用血浆非靶代谢组学探究制何首乌改善小鼠肾虚的作用机制。将35只ICR小鼠随机分为空白组(C)、模型组(M)以及黑豆汁炖24 h (braising with black bean sauce for 24 h,BB24 h)、32 h (BB32 h)、40 h (BB40 h)制何首乌给药组。造模后连续给药14天,取血检测生化指标,并通过超高效液相-线性离子阱-轨道阱串联质谱(UPLC-LTQ-orbitrap XL)测定各组小鼠血浆中内源性代谢物的变化,筛选差异代谢物。实验结果显示,与模型组相比,黑豆汁炖24 h给药组中雌激素(estradiol,E2)水平显著升高(P<0.05);黑豆汁炖32 h给药组中皮质醇(cortisol,Cort)水平显著升高(P<0.05),制何首乌对肾虚小鼠的激素水平有明显改善。制何首乌干预治疗后,小鼠血浆中共发现70个差异代谢物,主要涉及甘油磷脂代谢、花生四烯酸代谢、苯丙氨酸代谢、苯丙氨酸、酪氨酸和色氨酸的生物合成、亚油酸代谢等5条通路。研究提示制何首乌可能是通过改善小鼠下丘脑—垂体—肾上腺轴的紊乱并调节脂质代谢的水平,从而发挥补肝肾的作用。血浆中的差异代谢物与化学成分群的相关性分析发现二苯乙烯苷类及糖类成分可能是制何首乌发挥补肾作用的药效物质基础。本文为揭示制何首乌发挥补肾作用的潜在药效物质基础及补肝肾作用机制研究提供了新的参考和依据。本研究涉及的动物实验操作均遵循中国中医科学院中药研究所实验动物福利伦理委员会的规定并通过动物实验伦理审查[批准号:SYXK (京) 2019-0003]。

Plasma nontargeted metabolomics technology was developed for investigating the effect and mechanism of improving kidney deficient in mice of Polygoni Multiflori Radix Praeparata.Thirty-five ICR mice were randomly divided into the control group,the model group,the BB24 h(braising with black bean sauce for 24 hours)group,the BB32 h group,and the BB40 h group.Biochemical indices in blood plasma of mice were measured by collecting eye blood after modeling.Changes in plasma endogenous metabolites of mice from each group were determined by ultra-performance liquid chromatography-linear trap quadrupole-orbitrap XL(UPLC-LTQ-orbitrap XL),and differential metabolites were screened.The results of pharmacodynamic investigation showed that compared to the model group,the levels of estradiol increased obviously in the BB24 h(P<0.05),and the levels of cortisol increased obviously in BB32 h(P<0.05).The hormone level of mice with kidney deficiency was significantly improved after taking processed Polygonum multiflorum.A total of 70 differential endogenous metabolites in blood plasma of mice were identified from all treatment groups,which mainly involved glycerophospholipid meta-bolism,arachidonic acid metabolism,phenylalanine metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,and linoleic acid metabolism.The study indicated that Polygoni Multiflori Radix Praeparata may play the role of tonifying liver and kidney by improving the disorder of hypothalamic-pituitary-adrenal axis and regulating lipid metabolism in mice.Correlation analysis on differential metabolites in blood plasma and the chemical constituents showed that stilbene glycosides and saccharides may be the key pharmacodynamic material basis.The present study provides a new reference and theoretical foundation for revealing the potential pharmacodynamic material basis and mechanism investigation on tonifying liver and kidney of Polygoni Multiflori Radix Praeparata.This study was carried out following the ethical guidelines and regulations for the use of laboratory animals of the Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences and passed the animal experimental ethical review[No.SYXK(Jing)2019-0003].