基于氨基酸代谢研究白芍-甘草药对抗缺血性脑卒中的作用机制

To study the mechanism of action of the drug pair Radix Paeoniae Alba and Licorice against ischemic stroke based on amino acid metabolism

目的:从氨基酸靶向代谢角度,探讨"白芍-甘草"抗缺血性脑卒中的作用机制及其对mGluR1和GABAB的表达影响。方法:采用大鼠神经mNSS评分进行神经功能评价;超高效液相色谱-四级杆-飞行时间串联质谱(ultra performance liquid chromatography-quadrupole time of flight-mass spectrometry,UPLC-QTOF-MS)分析技术,建立大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)大鼠脑组织氨基酸代谢谱;比较分析模型组、假手术组、给药组,筛选出具有显著性差异的内源性氨基酸代谢物,采用qRT-PCR法检测大鼠脑组织的GABA-B受体mRNA表达和mGluR受体mRNA表达。采用Western blot法检测大鼠脑组织的GABA-B受体和mGluR受体表达。结果:与模型组比较,白芍-甘草给药后,大鼠神经功能损伤评分显著性降低;氨基酸靶向代谢组学结果表明,31种氨基酸及其衍生物可能参与缺血性脑卒中的发病与治疗过程;白芍-甘草给药后,MCAO大鼠GABABR1 mRNA、GABAB、mGluR1 mRNA和mGluR1总蛋白表达水平均有显著回调(P<0.01或P<0.05)。结论:"白芍-甘草"抗缺血性脑卒中的作用机制可能与氨基酸代谢通路密切相关,特别是与脑内牛磺酸、丝氨酸、谷氨酰胺等的含量变化有关,为缺血性脑卒中提供了一种新的研究切入点。

OBJECTIVE To investigate the anti-ischemic stroke mechanism of"Radix Paeoniae Alba and Licorice"and its effect on the expression of mGluR1 and GABAB from the perspective of targeted amino acid metabolism.METHODS mNSS score was used to evaluate nerve function.UPLC-QTOF-MS technique was used to establish amino acid metabolism profile in brain tissue of MCAO rats.The model group,sham operation group and drug administration group were compared and analyzed,and the endogenous amino acid metabolites with significant differences were screened out.qRT-PCR was used to detect the mRNA expression of GABA-B receptor and mGluR receptor in rat brain tissue.GABA-B receptor and mGluR receptor expression in rat brain were detected by Western blot.RESULTS Compared with model group,after administration of Radix Paeoniae Alba and Licorice,the neurological function injury score of rats was significantly decreased;Amino acid targeted metabolomics showed that 31 amino acids and their derivatives may be involved in the pathogenesis and treatment of ischemic stroke.The expression levels of GABABR1 mRNA,GABAB,mGluR1 mRNA and total protein of mGluR1 in MCAO rats were significantly reduced after administration of Radix Paeoniae Alba and Licorice.CONCLUSION The anti-ischemic stroke mechanism of"Radix Paeoniae Alba and Licorice"may be closely related to amino acid metabolic pathway,especially the contents of taurine,serine,glutamine,etc.,which provide a new research entry point for ischemic stroke.