通塞脉微丸干预缺血性脑中风大鼠的脑组织代谢组学研究

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

目的研究缺血性中风模型大鼠脑组织代谢的变化特点和通塞脉微丸对异常代谢的调节作用,寻找通塞脉微丸治疗缺血性脑中风可能的代谢生物标志物,以探索通塞脉微丸治疗缺血性脑中风的作用机制。方法采用电凝法制造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,运用气相色谱-质谱(GC-MS)技术,对各组脑组织中内源性代谢物进行测定,采用偏最小二乘法-判别分析(PLS-DA)对多变量数据进行分析,t检验方法进行显著性统计分析。结果模式识别中模型组大鼠能较好地与其他3组分开。与假手术组大鼠相比,模型组大鼠脑组织中的丙氨酸、丝氨酸、甲硫氨酸、柠檬酸、组氨酸、花生四烯酸(AA)和二十二碳六烯酸(DHA)的含量升高,而核糖、6-磷酸葡萄糖和肌苷的含量下降,提示脑组织中这10种内源性物质为脑缺血的潜在生物标志物。这些内源性物质在通塞脉组大鼠体内得到了不同程度的恢复,而花生四烯酸和二十二碳六烯酸在通塞脉组与模型组大鼠体内的含量有明显差异,说明通塞脉微丸对花生四烯酸和二十二碳六烯酸这两种潜在生物标志物有明显的调节作用。结论通塞脉微丸可以使造模后大鼠的体内代谢物含量有不同程度的恢复,提示其治疗作用可能与相关代谢通路的调节有关。

Aim To observe cerebral metabolic changes of ischemic cerebral apoplexy rat and the regulating action of Tongsaimai pellets on abnormal metabolism,and to find the potential biomarkers.To explore mechanisms of Tongsaimai pellets in treatment of ischemic cerebral apoplexy.Methods 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.Endogenous metabolites of four groups were determined with GC-MS.Partial least squares discriminant analysis（PLS-DA） was applied to analyze multivariate data,and T-test was used in significant statistical analysis.Results In scatter plot of rat brain samples,model group could be distinguished from 3 other groups.Compared with sham-operation group rats,alanine,serine,methionine,citric acid,histidine,arachidonic acid,docosahexaenoic acid increased in model group rats,while ribose,glucose-6-phosphate and inosine decreased,so these metabolites were potential metabolic biomarkers.These endogenous metabolites got restoration in Tongsaimai group rats.Arachidonic acid（AA） and docosahexaenoic acid（DHA） were statistically different between Tongsaimai pellets group and model group rats,showing that Tongsaimai pellets could obviously regulate AA and DHA.Conclusion Changed metabolities can be certainly recovered by Tongsaimai pellets,and the treatment of Tongsaimai pellets can be connected with the regulation of related metabolic pathways.