基于代谢组学分析电针对癫痫大鼠血浆代谢物变化的影响

Effects of EA on Plasma Metabolites in Epileptic Rats Based on Metabolomics Analysis

目的:应用代谢组学技术研究电针对癫痫模型大鼠的血浆代谢模式,初步探讨电针治疗癫痫的生物标志物,揭示电针改善癫痫的潜在代谢调节机制。方法:实验大鼠分为3组,即空白组、模型组和电针组,每组10只SD大鼠,通过戊四氮腹腔注射的方法制备癫痫模型。电针组取“百会”“大椎”穴,15 min/次,1次/d,连续28 d。评价模型组与电针组大鼠癫痫发作情况;HE染色法观察各组大鼠海马病理改变;代谢组学技术分析各组大鼠的血浆代谢轮廓,经PCA和OPLS-DA等多元统计方法分析,结合数据库筛选组间差异代谢物,差异代谢物经KEGG富集代谢通路。结果:与空白组比较,模型组大鼠癫痫发作频率升高显著(P<0.01),逃避潜伏期显著延长,60 s内穿越平台次数及平台象限停留时间减少,海马神经元损伤明显,共筛选出11个显著差异代谢物,KEGG富集分析到4条显著差异代谢通路。与模型组比较,电针组大鼠癫痫发作频率显著降低(P<0.01),逃避潜伏期显著缩短,60 s内穿越平台次数及平台象限停留时间延长,海马神经元损伤减轻,共筛选出显著差异代谢物7个,富集分析得到差异代谢通路2条,均参与了色氨酸代谢。结论:电针可降低癫痫大鼠的发作频率、改善海马结构损伤,其生物学机制涉及调节脂质代谢、氨基酸代谢和能量代谢等相关途径。

Objective:To study the effects of electro-acupuncture(EA)on plasma metabolite patterns in epileptic model rats by applying metabolomics techniques,to preliminarily investigate biomarkers for EA treatment of epilepsy and to reveal potential metabolic regulation mechanisms.Methods:30 SD rats were randomly divided into the blank group,the model group and the EA group,with 10 rats in each group.The epilepsy model was prepared by intraperitoneal injection of Pentylenetetrazol(PTZ).The EA group was treated with electro-needling Baihui(DU20)and Dazhui(DU14),15 min each time,once a day for 28 consecutive days.The epileptic seizures were evaluated in the model group and the EA group.The pathological changes of hippocampus were observed by HE staining.Metabolomics techniques were employed to analyze the plasma metabolic profiles of each group of rats.Multivariate statistical methods such as PCA and OPLS-DA were used for analysis.Differential metabolites between groups were identified by combining with database screening.Differential metabolites were then subjected to KEGG enrichment analysis to identify metabolic pathways.Results:Compared with those in the blank group,the seizure frequency was significantly increased(P<0.01),the escape latency was significantly prolonged,the frequency of crossing the platform and the residence time in the platform quadrant were reduced within 60 s,and the damage of hippocampal neurons was obvious in the model group.A total of 11 metabolites with significant differences were selected,and 4 metabolic pathways with significant differences were identified by KEGG enrichment analysis in the model group.Compared with those in the model group,the seizure frequency was significantly reduced(P<0.01),the escape latency was significantly shortened,the frequency of crossing the platform and the staying time of the platform quadrant were prolonged within 60 s,and the damage of hippocampal neurons was reduced in the EA group.A total of 7 significantly different metabolites were selected,and 2 different metabolic pathways were obtained through enrichment analysis,all of which were involved in tryptophan metabolism in the EA group.Conclusion:EA treatment can reduce seizure frequency and improve hippocampal structural damage in epileptic rats,and its biological mechanism involves regulating lipid metabolism,amino acid metabolism,energy metabolism and other related pathways.