基于miRNA组学探讨补肾调冲汤对薄型子宫内膜大鼠作用机制

Study on the Mechanism of Bushen Tiaochong Decoction(补肾调冲汤)on Thin Endometrial Rats Based on Mi RNA Omics

目的运用高通量测序技术筛选补肾调冲汤干预薄型子宫内膜大鼠模型中差异表达的miRNA,并对其进行生物信息学分析,揭示补肾调冲汤对薄型子宫内膜大鼠的作用机制。方法制备薄型子宫内膜大鼠模型,使用补肾调冲汤干预,以生理盐水做阴性对照,取两组大鼠子宫内膜组织进行总RNA提取、RNA质检、文库构建后,使用测序平台为Illumina HiSeq2500进行高通量测序,以P<0.05的标准筛选差异miRNA,利用miRanda和RNAhybrid得出差异表达miRNA候选靶基因,进行基因本体论(GO)、京都基因与基因组百科全书(KEGG)富集分析。结果共有48个差异miRNA,其中包括19个上调,29个下调,共得出2124个靶基因,其中涉及889个生物过程、146个细胞成分、163个分子功能,KEGG信号通路有AMPK信号通路、Ras信号通路、mTOR信号通路、NF-κB信号通路等。结论补肾调冲汤可能通过调节rno-miR-292-3p、rno-miR-292-5p、rno-miR-291a-3p、rno-miR-291b、rno-let-7c-1-3p、rno-miR-383-5p、rno-miR-338-5p等miRNA表达作用于薄型子宫内膜模型大鼠,可能涉及调控AMPK信号通路、Ras信号通路、mTOR信号通路、NF-κB信号通路等参与薄型子宫内膜的子宫内膜增殖、子宫内膜容受性建立及胚胎植入等重要过程。

Objective To use high-throughput sequencing technology to screen the differentially expressed miRNAs of Bushen Tiaochong Decoction(补肾调冲汤)for intervention in a thin endometrial rat model,and to conduct bioinformatics analysis on them,and reveal the mechanism of Bushen Tiaochong Decoction on thin endometrial rats.Methods Prepare a thin endometrial rat model,use Bushen Tiaochong Decoction as a negative control,take two groups of rat endometrial tissues for total RNA extraction,RNA quality inspection,library construction,and use the sequencing platform perform high-throughput sequencing for Illumina HiSeqTM2500,screen differential miRNAs with a standard of P<0.05,use miRanda and RNAhybrid to obtain differentially expressed miRNA candidate target genes,and perform GO and KEGG enrichment analysis.Results A total of 48 differential miRNAs,including 19 up-regulated and 29 down-regulated,yielded a total of 2124 target genes,which involved 889 biological processes,146 cell components,and 163 molecular functions.The KEGG signaling pathway had AMPK signaling pathway,Ras signaling pathway,m TOR signaling pathway,NF-κB signaling pathway,etc.Conclusion Bushen Tiaochong Decoction may regulate rno-miR-292-3p,rno-miR-292-5 p,rno-miR-291a-3p,rno-miR-291b,rno-let-7c-1-3p,rno-miR-383-5p,rno-miR-338-5p and other miRNA expressions affect thin endometrial model rats,which may involve the regulation of AMPK signaling pathway,Ras signaling pathway,m TOR signaling pathway,NF-κB signaling pathway and other signaling pathways.The thin endometrium is an important process such as endometrial proliferation,endometrial receptivity establishment and embryo implantation.