当归挥发油给药大鼠尿液比较代谢组学研究

Comparative metabonomics study on urine in rat treated by Angelica sinensis volatile oil

为了分析当归挥发油(Angelica sinensis volatile oil,ASVO)对正常大鼠尿液代谢产物的影响,揭示ASVO影响大鼠体内代谢的可能途径。该实验将50只雄性Wister大鼠随机分成空白对照组、ASVO高(0.352 mL·kg-1)、中(0.176 mL·kg-1)、低剂量(0.088 mL·kg-1)组和阿司匹林(aspirin,ASP)对照组,每组10只,分别给予生理盐水和高、中、低剂量的ASVO及ASP各0.2 mL,连续给药3 d,并利用大鼠代谢笼收集各组大鼠12,24,36,48 h的尿液,应用GC-MS检测不同时段的大鼠尿液代谢指纹图谱,采用主成分分析(principal component analysis,PCA)方法和正交偏最小二乘分析方法(orthogonal partial leastsquares discriminant analysis,OPLS-DA)分析,通过变量重要性投影(vriable importance in the projection,VIP)和T检验筛选潜在生物标志物。结果显示,ASVO中剂量组在36 h时对大鼠尿液代谢物影响最大。与空白对照组相比,ASP组与ASVO给药组大鼠尿液中7种代谢物有显著变化(P<0.05),其中乌头酸、琥珀酸、柠檬酸、α-酮戊二酸、甘氨酸、苹果酸的含量呈升高趋势(P<0.05),前列腺素含量呈下降状态(P<0.01),认为ASVO与ASP的作用机制具有相似性。它们对体内代谢的影响可能主要集中于能量代谢、氨基酸代谢以及脂质代谢途径。大鼠灌胃ASVO后可提高机体的能量代谢,抑制炎性物质的产生、增强机体的免疫能力,该结果为进一步阐释ASVO药理作用提供了依据。

Metabonomics was employed to investigate the effect of Angelica sinensis volatile oil（ASVO）to the endogenous metabolites of normal rats, and to reveal the possible ways of metabolism in rats caused by ASVO. The fifty male Waster rats were randomly divided into five groups（each consists of 10 rats）, such as control group, high dose group of ASVO, middle dose group of ASVO, low dose group of ASVO, and Aspirin group. They were given 0.9% saline, 0.352 mL·kg-1 ASVO, 0.176 mL·kg-1 ASVO, 0.088 mL·kg-1 ASVO and ASP respectively with the equal volume of 0.2 mL. Drugs and vehicle were given for 3 successive days. The urine was collected at 12, 24, 36, 48 h after modeling with metabolic cages. Rat urine metabolic fingerprint in different stages was analyzed using GC-MS, based on which the principal component analysis（PCA）and orthogonal partial least-squares discriminant analysis（OPLS-DA）models were established for metabonomic analysis. Potential biomarkers were screened by using variable importance in the projection（VIP）and T test. It was revealed that the middle dose of ASVO at 36 h induces a substantial change in rat urine. Compared with control group, seven kinds of endogenous metabolites in ASP group and ASVO group change significantly（P〈0.05）, among which aconitic acid, succinic acid, citric acid, α-ketone glutaric acid, glycine and malic acid content had an upward trend（P〈0.05）and prostaglandin content had a downward trend（P〈0.01）. The mechanism of ASVO and ASP have the similarity. It is likely that ASVO intervenes the metabolic process by affecting the energy, amino acid and lipid metabolism. Our work also indicates that rats administrated with ASVO can increase the energy metabolism of the body, induce the production of inflammatory substances and strengthen the body＇s immune ability. The result has also provide a proof for futher interpret ASVO pharmacological effects.