基于UPLC-MS探讨痰湿壅盛型高血压病小鼠血浆代谢组学特点

Study on the characteristics of plasma metabolomics in murine model of hypertension with excessive accumulation of phlegm-dampness based on UPLC-MS

目的:通过检测痰湿壅盛型高血压病模型小鼠的血浆代谢组学变化,分析挖掘该证候的潜在生物学标志物,从整体角度阐释痰湿壅盛型高血压病模型小鼠的代谢特征。方法:高脂饲料喂养C57BL/6Cnc小鼠25周,观察小鼠一般情况和主动脉组织形态,UPLC-MS分析血浆代谢组变化,结合多维统计和单位统计分析筛选组间差异代谢物,并进行KEGG代谢通路富集分析。结果:模型组小鼠出现痰湿壅盛证特征表现,体质量、Lee’s指数、收缩压、舒张压、血糖/胰岛素、瘦素水平较对照组显著升高(*P<0.05),空腹胰岛素显著降低(P<0.05),主动脉出现明显的形态改变和脂质沉积。UPLC-MS共筛选出差异代谢物125个(其中64个升高,61个降低;VIP>1且P<0.05),差异最大的代谢物依次为磷脂酰胆碱、溶血磷脂酰胆碱、L-异亮氨酸、乙酰肉碱等。差异代谢物主要富集于胆碱代谢、GnRH信号通路、氨基酸的合成、FcγR介导的吞噬作用、磷脂酶D信号途径等(P<0.01),以及产热、EGFR信号通路、NF-κB信号通路等(P<0.05)。结论:磷脂酰胆碱、溶血磷脂酰胆碱、L-异亮氨酸、乙酰肉碱可作为痰湿壅盛型高血压病小鼠模型的特征血浆代谢标志物。

Objective: To explore the syndrome of internal exuberance of phlegm-damp markers by assaying the plasma metabonomics in the murine model of hypertension and explain the metabolic characteristics of the murine model of hypertension with syndrome of internal exuberance of phlegm-damp from the overall perspective. Methods: C57 BL/6 Cnc mice were fed with high fat diet for 25 weeks to induce the murine model of hypertension with syndrome of internal exuberance of phlegmdamp. The general status of mice and the tissue morphology of aorta were observed. The plasma metabonomics were assayed by UPLC-MS, the differential metabolites between groups were screened by multi-dimensional statistics and unit statistical analysis, and the enrichment of KEGG metabolic pathway was analyzed. Results: Model mice demonstrated the characteristics of syndrome of internal exuberance of phlegm-damp, the body weight, Lee’s index, systolic blood pressure, diastolic blood pressure, blood glucose/insulin and leptin levels were significantly increased than the control group(P<0.05), while fasting insulin decreased significantly(P<0.05). Obvious morphological changes and lipid deposition in the aorta were observed in model mice. A total of 125 different metabolites(64 increased and 61 decreased;VIP>1 and P<0.05) were screened out by UPLCMS. The most significant metabolites were phosphatidylcholine, lysophosphatidylcholine, L-isoleucine and acetylcarnitine,etc.. The main metabolites were choline metabolism, GnRH signaling pathway, amino acid biosynthesis and FCγR-mediated phagocytosis(P<0.01), and thermogenesis, EGFR signaling pathway, NF-κB signaling pathway, etc.(P<0.05). Conclusion: Phosphatidylcholine, lysophosphatidylcholine, L-isoleucine and acetylcarnitine can be used as the characteristic plasma metabolic markers in the model of hypertension with syndrome of internal exuberance of phlegm-damp.