基于UHPLC-QE-MS的银星养脑片治疗血管痴呆性大鼠的血清代谢组学研究

Effects of Yinxing Yangnao Tablet(银星养脑片)in Treatment of Vascular Dementia in Rats Based on UHPLC-QE-MS by Serum Metabolomics

目的:基于超高效液相色谱-质谱联用(UHPLC-QE-MS)技术探讨银星养脑片对血管痴呆性大鼠血清中内源性标志物的影响,并初步探讨其发挥药效的作用通路。方法:采用双侧颈总动脉结扎法诱导血管性痴呆模型,将SD大鼠随机分为正常对照组、模型对照组、吡拉西坦片0.4 g/kg组、银星养脑片0.27 g/kg组,每组10只。经双侧颈总动脉结扎后14 d开始给药,连续给药14 d,给药结束后立即进行水迷宫测试并将动物脑组织进行HE染色,应用UHPLC-QE-MS技术进行血清样品的分离和数据采集,采用主成分分析法(PCA)和正交偏最小二乘判别分析法(OPLS-DA)筛选出相关差异代谢物,并对已经鉴定的代谢标志物使用KEGG数据库进行可能的代谢通路分析。结果:水迷宫结果显示,与正常对照组相比,模型对照组动物出现明显认知功能障碍;HE染色证明模型对照组大鼠海马CA1区神经元出现病理性损伤,银星养脑片0.27 g/kg组有一定的改善;PCA结果显示,正常对照组、模型对照组和银星养脑片组大鼠血清代谢物发生了明显的变化;通过OPLS-DA分析最终确定在3个组中的40个显著差异代谢物;通路分析显示,主要与支链氨基酸生物合成代谢途径有关。结论:银星养脑片对血管性痴呆大鼠的治疗作用可能与支链氨基酸合成代谢有关。

Objective:To investigate the effects and the underlying mechanism of Yinxing Yangnao Tablet on the potential endogenous markers of vascular dementia rats by UHPLC-QE-MS,and explore its effective pathways.Methods:The vascular dementia model was induced by permanent ligation of bilateral common carotid arteries.SD rats were randomly divided into a control group,a model group,a piracetam group(0.4 g/kg),and a Yinxing Yangnao Tablet group(0.27 g/kg),with 10 rats in each group.Fourteen days after ligation of bilateral common carotid arteries,drugs were orally administered for 14 days.After the last administration,the Morris water maze test was performed.Rat brain tissues were stained by HE.UHPLC-QE-MS technology was used for serum sample separation and data were collected.Principal component analysis(PCA)and orthogonal projections to latent structures discriminant analysis(OPLS-DA)were used to screen related differential metabolites,and identified metabolomic biomarkers were subjected to metabolomic pathway analysis by KEGG.Results:The Morris water maze test showed that compared with the control group,the model group showed obvious cognitive disorder.HE staining showed obvious neuronal damage in the hippocampal CA1 region in the model group,while the damage was significantly improved in the Yinxing Yangnao Tablet group.PCA results showed that the serum metabolites were significantly changed in the control group,the model group,and the Yinxing Yangnao Tablet group.Forty significant biomarkers were obtained in three groups by OPLS-DA.The metabolic pathway analysis showed that the therapeutic effect of Yinxing Yangnao Tablet was achieved through the metabolic pathways related to the biosynthesis of branched-chain amino acids.Conclusion:This study preliminarily clarifies the therapeutic mechanism of Yinxing Yangnao Tablet against vascular dementia,which may be related to the biosynthesis and metabolism of branched-chain amino acids.