通塞脉微丸干预缺血性中风模型大鼠的血浆代谢组学研究

Plasma metabonomics study of ischemic cerebral apoplexy rats treated with Tongsaimai pellets

目的:通过分析缺血性中风模型大鼠血浆内小分子的变化,研究缺血性中风导致的代谢异常变化及通塞脉微丸对异常代谢的调节作用,寻找缺血性脑中风模型可能的代谢生物标志物,探索通塞脉微丸的代谢性作用机制。方法:采用电凝法造成大鼠缺血性中风模型,将大鼠分为模型组、假手术组、通塞脉组和阳性药组,每组8只。通塞脉组按生药量13.2 g.kg-1.d-1灌胃给药,阳性药组给予尼莫地平32 mg.kg-1.d-1,模型组、假手术组灌服等体积蒸馏水,连续给药7 d。采集模型组、假手术组血浆和通塞脉组、阳性药组大鼠给药第1,3,7天的血浆,采用气相色谱-质谱(GC-MS)对血浆中内源性分子进行测定,采用偏最小二乘法-判别分析(PLS-DA)对多变量数据进行分析并建立模型,t检验方法进行统计分析。结果:与假手术组大鼠相比,模型组大鼠血浆中的丙酮酸、牛磺酸、羟脯氨酸的含量明显升高,而乳酸、甘油酸、氨基丙二酸、果糖、色氨酸和亮氨酸显著下降,提示血浆中这9种内源性物质为脑缺血的潜在生物标志物。通塞脉微丸给药使得上述内源性物质(除牛磺酸)得到了不同程度的恢复。结论:通塞脉微丸可以使造模后大鼠血浆中异常变化的代谢物水平有不同程度的恢复,提示其药效作用可能与其对相关代谢途径的调节有关。

Objective： To observe abnormal metabolic changes caused by ischemic cerebral apoplexy and the regulating action of Tongsaimai pellets on abnormal metabolism by analyzing the change of small molecules in plasma of ischemic cerebral apoplexy rat.To find the potential biomarkers,and to explore metabolic mechanisms of Tongsaimai pellets.Method： Rat models of middle cerebral artery occlusion was established with electric coagulation,and rats were divided into 4 groups,model group,sham-operation group,Tongsaimai pellets group and positive control group.Tongsaimai pellets and positive control group were orally administrated by 13.2 g·kg-1·d-1 of crude drugs and 32 mg·kg-1·d-1 of Nimodipine respectively,model and sham-operation group by equal volume of distilled water for a week.Plasma of model and sham-operation group were collected,and plasma of Tongsaimai pellets and positive control group were collected on the 1st,3rd,7th day after administration.Endogenous metabolites of four groups were determined with GC-MS.Partial least squares discriminant analysis（PLS-DA） was applied to analyze multivariate data and set up model,and T-test was used in significant statistical analysis.Result： Compared with sham-operation group rats,pyruvic acid,taurine and hydroxyproline obviously increased in model group rats,while lactic acid,glyceric acid,aminomalonic acid,fructose,tryptophan and leucine significantly decreased,so these metabolites were potential metabolic biomarkers.These endogenous metabolites except taurine got restoration in Tongsaimai group rats.Conclusion： Abnormal metabolite level in plasma can be certainly recovered by Tongsaimai pellets,and the treatment of Tongsaimai pellets can be connected with the regulation of related metabolic pathways.