桃红四物汤对缺血性脑卒中损伤后微小RNA的影响

Effect of Taohong Siwu Decoction on MicroRNAs After Ischemic Stroke

目的研究桃红四物汤(Taohong Siwu Decoction,THSWD)对缺血性脑卒中的微小RNA(microRNA,miRNA)表达谱的影响,探讨THSWD治疗缺血性脑卒中的分子机制。方法建立大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)诱导的局灶性脑缺血大鼠模型,采用单细胞RNA测序技术检测THSWD治疗后MCAO大鼠miRNA表达谱。采用GO和KEGG富集分析方法分析miRNA作用于靶基因的功能。对表达差异显著的miRNA进行RT-qPCR实验验证。结果THSWD干预后,MCAO大鼠脑梗死区有13个miRNA表达发生变化,其中6个miRNA表达上调,7个miRNA表达下调。利用miRanda进行预测,确定了这13个miRNA的靶基因,构建了miRNA-mRNA网络,通过RT-qPCR验证了其中6个差异表达的miRNA(rno-miR-653-5p、rno-miR-216b-5p、rno-miR-3547、rno-miR-217-5p、rno-miR-758-3p和rno-miR-223-3p)。结论THSWD可以通过逆转某些miRNA的异常表达来治疗缺血性脑卒中。

Objective To investigate the effect of Taohong Siwu Decoction(THSWD)on the expression profile of microRNAs(miRNAs)in improving ischemic stroke,as well as the potential effect and molecular mechanism of THSWD in protecting against ischemic stroke.Methods A rat model of focal cerebral ischemia induced by middle cerebral artery occlusion(MCAO)was established,and the single-cell RNA sequencing technique was used to measure the expression profile of miRNAs in MCAO rats after THSWD treatment.Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were used to analyze the function of these miRNAs on target genes.RT-qPCR was used for the validation of differentially expressed miRNAs.Results After THSWD treatment,there were changes in 13 miRNAs in the cerebral infarction area of MCAO rats,among which 6 were upregulated and 7 were downregulated.The target genes of these 13 miRNAs were identified by miRanda,and a miRNA-mRNA network was constructed.Six differentially expressed miRNAs were verified by RT-qPCR,i.e.,rno-miR-653-5p,rno-miR-216b-5p,rno-miR-3547,rno-miR-217-5p,rno-miR-758-3p,and rno-miR-223-3p.Conclusion THSWD can improve stroke by reversing the abnormal expression of some miRNAs.