电针对大脑中动脉栓塞大鼠缺血侧大脑皮层miRNA表达谱的影响

Effect of electroacupuncture on miRNA expression profile in ischemic cerebral cortex of rats with middle cerebral artery embolization

目的:观察miRNA在脑梗死大鼠和电针治疗后的大鼠大脑皮层组织中的表达差异,寻找与脑梗死相关的潜在治疗靶点。方法:将18只SD大鼠随机分为正常组、模型组和电针组,每组6只。模型组与电针组采用Longa改良线栓法制备右侧大脑中动脉闭塞(MCAO)模型。电针组于造模后电针双侧“足三里”“内关”穴,连续7 d。采用苏木精-伊红(HE)染色观察各组大鼠脑组织病理学变化,并对各组缺血侧大脑皮层进行转录组测序,筛选差异表达的miRNA并对其靶基因进行GO功能富集分析和KEGG通路富集分析。结果:电针组大鼠脑组织病理损伤较模型组明显减轻。与模型组相比,正常组差异表达的miRNA共401个,其中340个上调,61个下调。与电针组相比,模型组差异表达的miRNA共21个,其中9个上调,12个下调。MCAO病理状态下,miR-743a-3p、miR-11980-z、miR-871-3p、miR-743b-3p、miR-741-3p、miR-6715-x、novel-m0082-3p表达下调,miR-7-z表达上调(P<0.05),这些miRNA的表达均在电针干预后逆转。代谢通路、MAPK信号通路、Wnt信号通路等与神经系统相关的功能和通路被富集。结论:电针治疗后,脑梗死相关的miRNA表达谱发生了显著的变化;电针可能通过调控代谢通路、MAPK信号通路、Wnt信号通路等在治疗脑梗死过程中发挥保护作用。

Objective:To explore the difference of miRNA expression in cerebral cortex of rats with cerebral infarction and electroacupuncture group to find potential targets related to the treatment of cerebral infarction.Methods:Eighteen SD rats were randomly divided into normal group,model group and electroacupuncture group,with 6 rats in each group.The model of right middle cerebral artery occlusion was established by Longa modified thread occlusion method in model group and electroacupuncture group.After the establishment of the model,the electroacupuncture group was electroacupuncture at bilateral"Zusanli"and"Neiguan"points for 7 consecutive days.The pathological changes of MCAO rats treated with electroacupuncture were observed by hematoxylin-eosin staining,and the transcriptional sequences of cerebral cortex on the ischemic side of each group were sequenced.The differentially expressed miRNAs were screened and their target genes were analyzed by GO function enrichment analysis and KEGG pathway enrichment analysis.Results:The pathological results showed that electroacupuncture significantly reduced the pathological injury in MCAO rats.Compared with the model group,the number of differentially expressed miRNAs in the normal group was 401,among which 340 were up-regulated and 61 were down-regulated.Compared with the electroacupuncture group,there were 21 differentially expressed miRNAs,of which9 were up-regulated and 12 were down-regulated in the model group,and under the pathological condition of MCAO,miR-743a-3p,miR-11980-z,miR-871-3p,miR-743b-3p,miR-741-3p,miR-6715-x,novel-m0082-3p with down-regulated expression and miR-7-z with up-regulated expression were reversed after electroacupuncture intervention.The results of GO and KEGG enrichment analysis of differential miRNA source genes showed that Metabolic pathway,MAPK signaling pathway and Wnt signaling pathway and other nervous system-related functions and pathways were enriched.Conclusion:After the intervention of electroacupuncture,the miRNA expression profile associated with cerebral infarction has changed significantly.Electroacupuncture may play a role in the treatment of cerebral infarction by regulating Metabolic pathway,MAPK signal pathway,Wnt signal pathway and so on.