分析当归(Angelica sinensis,AS)不同炮制品挥发油对正常大鼠尿液代谢产物的影响,揭示其影响大鼠体内可能的代谢途径。该实验将50只雄性Wister大鼠随机分成空白对照组和当归炭挥发油(volatile oils from charred AS,C-ASVO)、酒当归挥发油(volatile oils from parched AS with alcohol,J-ASVO)、土当归挥发油(volatile oils from parched AS with soil,T-ASVO)、油当归挥发油(volatile oils from parched AS with oil,Y-ASVO)组,每组10只,正常对照组灌胃等体积的0.5%的聚氧乙烯脱水山梨醇单油酸酯(吐温-80),各当归炮制品挥发油组每次灌胃挥发油剂量为0.176 m L·kg-1,连续给药3 d,利用大鼠代谢笼收集各组大鼠48 h的尿液。采用气相色谱质谱联用技术(gas chromatography-mass spectrometer,GC-MS)对大鼠尿液代谢物进行检测,并采用主成分分析方法(principal component analysis,PCA),正交偏最小二乘分析方法(orthogonal partial least-squares discriminant analysis,OPLS-DA)进行多元统计分析,通过变量重要性投影(variable importance in the projection,VIP)和t检验来选择差异代谢物。利用metabolic pathway analysis(Met PA)数据库构建差异代谢物的代谢通路。结果显示,与空白对照组相比,当归不同炮制品挥发油给药组大鼠尿液中31种代谢物有显著变化(P<0.05),并且当归不同炮制品挥发油对正常大鼠尿液代谢产物的影响存在差异,其中以J-ASVO的影响程度略强于C-ASVO,T-ASVO和Y-ASVO。它们对体内代谢的影响可能主要集中于能量代谢、氨基酸代谢、脂质代谢以及糖代谢。该研究补充了当归不同炮制品挥发油药物代谢和作用机制,为其中药药理作用深层次的研究提供思路。
目的:比较当归不同炮制品挥发油对脂多糖(Lipopolysaccharide,LPS)致大鼠急性炎症的干预作用。方法:通过腹腔注射LPS(100μg/kg)建立大鼠急性炎症模型,通过血常规和血清炎性介质及细胞因子检测,结合肺脏和肝脏组织病理学观察,评价酒当归挥发油(volatile oils from parching AS with wine,J-VOAS)、当归炭挥发油(volatile oils from charred AS,C-VOAS)、生当归挥发油(volatile oils from raw AS,S-VOAS)的抗炎活性。结果:与正常对照组相比,模型组白细胞计数(white blood cell count,WBC)、中性粒细胞百分比(neutrophil percentage,NE)和血小板计数(platelet Count,PLT)显著升高(P<0.05),淋巴细胞百分比(lymphocytes percentage,LY)显著降低(P<0.05);模型组血清IL-1β、IL-6、NO、TNF-α含量极显著升高(P<0.001),IL-10含量极显著降低(P<0.01);经J-VOAS、CVOAS、S-VOAS干预后上述血常规指标和血清炎性因介质及细胞因子均显著回调(P<0.05,P<0.01,P<0.001)。组织病理学观察发现,模型组大鼠肺泡扩张,肺泡及间质中大量炎性细胞浸润,肝小叶弥散性坏死,肝索排列紊乱,部分肝细胞发生颗粒变性和水肿;经J-VOAS干预后,其病理变化明显改善。结论:当归不同炮制品挥发油对LPS所致大鼠急性炎症模型均有干预作用,其中J-VOAS干预作用最好,其次为C-VOAS和S-VOAS。
This study aimed at determining the effects of Angelica sinensis(AS) on urinary metabolites in blood deficiency mice and exploring its replenishing blood mechanism. Gas chromatography–mass spectrometry(GC-MS) was applied to detect metabolites in the urine samples in different collection periods. Principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used to investigate the differences in metabolic profiles among control group(CG), blood deficiency model group(MG), AS groups, and Colla Corii Asini group(CCAG). The potential biomarkers were identified based on the variable importance in the projection(VIP), T-test, and National Institute of Standards and Technology(NIST) and mass spectra library. The metabolites were analyzed using metabolomics pathway analysis(Met PA) to build the metabolic pathways. Our results indicated that, on the seventh day, the levels of glucose, lactic acid, pyruvic acid, alanine, acetoacetic acid, and citric acid changed significantly in blood deficiency mice. However, these metabolic deviations came to closer to normal levels after AS intervention. The reversing blood-deficiency mechanism of AS might involve regulating synthesis and degradation of ketone bodies, Pyruvate metabolism, TCA cycle, and Glycolysis / Gluconeogenesis. In conclusion, metabonomics is a robust and promising means for the identification of biomarkers and elucidation of the mechanisms of a disease, thereby highlighting its importance in drug discovery.
WANG TaoSUN Hong-GuoHUA Yong-LiLI Peng-LingWEI Yan-Ming
