桃红四物汤对大脑中动脉闭塞大鼠lncRNA表达的影响

Effects of Tao Hong Si Wu decoction on lncRNA expression in rats with occlusion of middle cerebral artery

目的研究中药复方桃红四物汤(Tao Hong Si Wu decoction,THSWD)治疗大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)大鼠长链非编码RNA(long non-coding RNA,lncRNA)的表达,并确定THSWD治疗MCAO大鼠可能的分子机制。方法从对照组、MCAO组和MCAO+THSWD组各获得3个大脑半球组织。采用RNA测序技术鉴定三组中的lncRNA基因表达。鉴定了THSWD调节的lncRNA基因,然后构建了THSWD调节的lncRNA-mRNA网络。通过MCODE插件鉴定lncRNA-mRNA网络的模块。基因本体(gene ontology,GO)和京都基因与基因组百科全书数据库(kyoto encyclopedia of genes and genomes,KEGG)用于分析富集的生物功能和信号通路。鉴定了THSWD调节的lncRNA的顺式和反式调控基因。采用逆转录实时定量聚合酶链式反应(RT-qPCR)验证lncRNA。分子对接用于验证lncRNA-mRNA网络靶点和通路相关蛋白结合能力。结果在MCAO大鼠中,THSWD共调节了302个lncRNA。生物信息学分析表明,一些核心lncRNA可能在THSWD治疗MCAO大鼠中发挥重要作用,此外,我们进一步发现THSWD可能也通过lncRNA-mRNA网络以及网络富集的补体和凝血级联反应等多通路治疗MCAO大鼠。分子对接结果表明,THSWD活性化合物没食子酸和苦杏仁苷与蛋白质靶点具有一定的结合能力。结论THSWD可以通过调节lncRNA保护MCAO大鼠脑损伤,为THSWD治疗缺血性中风提供了新见解。

Aim To screen and study the expression of long non-coding RNA(lncRNA)in rats with middle cerebral artery occlusion(MCAO)with MCAO treated with Tao Hong Si Wu decoction(THSWD)and determine the possible molecular mechanism of THSWD in treating MCAO rats.Methods Three cerebral hemisphere tissue were obtained from the control group,MCAO group and MCAO+THSWD group.RNA sequencing technology was used to identify lncRNA gene expression in the three groups.THSWD-regulated lncRNA genes were identified,and then a THSWD-regulated lncRNA-mRNA network was constructed.MCODE plug-in units were used to identify the modules of lncRNA-mRNA networks.Gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG)were used to analyze the enriched biological functions and signaling pathways.Cis-and trans-regulatory genes for THSWD-regulated lncRNAs were identified.Reverse transcription real-time quantitative polymerase chain reaction(RT-qPCR)was used to verify lncRNAs.Molecular docking was used to identify lncRNA-mRNA network targets and pathway-associated proteins.Results In MCAO rats,THSWD regulated a total of 302 lncRNAs.Bioinformatics analysis suggested that some core lncRNAs might play an important role in the treatment of MCAO rats with THSWD,and we further found that THSWD might also treat MCAO rats through multiple pathways such as lncRNA-mRNA network and network-enriched complement and coagulation cascades.The results of molecular docking showed that the active compounds gallic acid and amygdalin of THSWD had a certain binding ability to protein targets.Conclusions THSWD can protect the brain injury of MCAO rats through lncRNA,which may provide new insights for the treatment of ischemic stroke with THSWD.