基于非靶向代谢组学研究前额叶皮质参与电针促醒的相关作用机制  

Exploring Related Mechanism of Prefrontal Cortex Involved in EA Therapy for Promoting Consciousness Based on Non-Targeted Metabolomics

作　　者：林友益 何毅琪 欧阳玉洁 姚冬 吴心虹 LIN Youyi;HE Yiqi;OUYANG Yujie;YAO Dong;WU Xinhong(Xiamen Fifth Hospital,Xiamen 361101,China;Xiamen University,Xiamen 361102,China)

机构地区：[1]厦门市第五医院,福建厦门361101 [2]厦门大学,福建厦门361102

出　　处：《针灸临床杂志》2024年第12期55-62,共8页Journal of Clinical Acupuncture and Moxibustion

基　　金：厦门市医疗卫生指导性项目,编号:3502Z20224ZD1294。

摘　　要：目的:采用非靶向代谢组学研究电针对脑外伤昏迷大鼠前额叶皮质代谢物的影响,探讨前额叶皮质参与电针促醒的相关作用机制。方法:将18只SPF级健康SD大鼠随机分成空白组、对照组和电针组3组,每组各6只。对照组和电针组均采用经典“自由落体撞击法”制备脑外伤大鼠模型。造模成功后对电针组针刺“百会”“水沟”穴位后给予电针刺激15 min,对照组仅针刺。实验完成后取前额叶皮质组织,应用Western blot法检测前额叶皮质γ-氨基丁酸b受体、水通道蛋白-4(AQP-4)及丙二醛-5(MDA-5)蛋白的表达,并用一维核磁共振氢谱技术(^(1)H NMR)分析各组前额叶皮质的代谢物,筛选差异代谢物及分析代谢轮廓,并将差异代谢物进行京都基因与基因组百科全书代谢通路富集分析。结果:与空白组比较,对照组前额叶皮质AQP-4和MDA-5蛋白表达量显著上升(P<0.01),GABA b受体蛋白表达量显著上升(P<0.01)。与对照组比较,电针组MDA-5蛋白表达量显著下降(P<0.01),AQP-4蛋白表达量明显下降,差异有统计学意义(P<0.05);GABA b受体蛋白表达量下降,但差异无统计学意义(P<0.05)。差异代谢物分析显示:对照组与空白组比较,共筛选出30种内源性差异代谢产物,两组间代谢物分布呈现明显离散,表明脑外伤大鼠造模成功后,脑组织内各代谢物的含量发生明显改变;电针组与对照组比较,大鼠脑组织代谢物轮廓分离明显,表明电针干预对代谢物产生显著影响;代谢通路富集分析显示差异代谢物主要富集通路为氨基酸代谢。结论:前额叶皮质参与电针促醒的分子机制可能与电针下调GABA b受体的表达及通过影响兴奋性氨基酸的代谢有关。Objective:To study the effect of electro-acupuncture(EA)on the metabolites of prefrontal cortex in coma rats with traumatic brain injury(TBI)based on non-targeted metabolomics,and to explore the related mechanism of prefrontal cortex participating in EA therapy for promoting consciousness.Methods:18 SPF healthy SD rats were randomly divided into the blank group,the control group and the EA group,with 6 rats in each group.The rat model of brain injury was prepared by the classic‘free fall impact method’.After successful modeling,the EA group was intervened with electro-needling Baihui(DU20)and Shuigou(DU26)for 15 min,and the control group was intervened by acupuncture.The prefrontal cortex tissue was taken,and the protein expressions ofγ-aminobutyric acid b receptor,aquaporin-4(AQP-4)and malondialdehyde-5(MDA-5)in the prefrontal cortex were detected by Western blot.The metabolites of the prefrontal cortex of each group were analyzed by 1H NMR(1H Nuclear Magnetic Resonance),the differential metabolites were subjected to Kyoto Encyclopedia of Genes and Genomes metabolic pathway enrichment analysis.Results:Compared with those in the blank group,the protein expressions of AQP-4,MDA-5 and GABA b receptor were significantly increased in the control group(P<0.01).Compared with those in the control group,the protein expressions of MDA-5 and AQP-4 were significantly decreased in the EA group(P<0.01,P<0.05);the protein expression of GABA b receptor was decreased,but the difference was not statistically significant.The analysis of differential metabolites showed that a total of 30 endogenous differential metabolites were screened out in the control group compared with those in the blank group.The distribution of the metabolites between the two groups showed obvious dispersion,indicating that the content of each metabolite in the brain tissue was significantly changed after successful modeling.Compared with those in the control group,the outline of metabolites in rat brain tissue was significantly separated in the EA gro

关 键 词：颅脑损伤 电针 前额叶皮质 代谢组学 兴奋性氨基酸 

分 类 号：R245.9[医药卫生—针灸推拿学]