Study On The Mechanism Of Bushen Tiaochong Decoction On Thin Endometrial Rats Based On MiRNAomics

OBJECTIVE:In this study,a thin endometrial rat model was used as the research object.The Bushen Tiaochong Decoction was used for intervention,and the high-throughput sequencing technology Illumina HiSeqTM M2500 sequencing platform was used to establish a thin endometrial rat model,Bushen Tiaochong Decoction and valeric acid After the intervention of estradiol,the endometrial miRNA expression profile of a thin endometrial rat model was screened for differential miRNAs and their target genes were predicted.GO and KEGG enrichment analysis were performed to screen out the biological processes,cellular components,molecular functions and Related signal pathways provide a scientific and objective theoretical basis for the clinical application of Bushen Tiaochong Decoction in the treatment of thin endometrium.Methods:(1)Using Bushen Tiaochong Decoction as the research prescription,42 unmated female(SPF grade)SD rats,each weighing 200±20g,were randomly selected according to the experimental requirements,and 2 estrus rats were randomly selected for thin uterus Preparation and verification of the endometrial model.Then the remaining 40 estrus rats were randomly divided into normal control group,model group,traditional Chinese medicine group and western medicine group,each with 10 rats,and the model group,traditional Chinese medicine group and western medicine group were prepared.Into a thin endometrial model rat.(2)The normal control group and the model group were given normal saline(1ml/100g)equal to their body weight,the traditional Chinese medicine group was given a crude drug quantity of 3.410g/ml Bushen Tiaochong Decoction,and the western medicine group was given estradiol valerate(0.1 mg/(kg·d))dissolved in(1ml/100g)of normal saline for intragastric administration,once a day,for 3 consecutive estrus cycles.(3)Subsequently,the rat endometrial tissues of each group were taken for Total RNA extraction,RNA quality inspection,and library construction.After the library inspection is qualified,Jikai Gene Co.,Ltd.is entrusted to complete high-throughput sequencing,and the sequencing platform is Illumina HiSeqTM2500.(4)Process the original sequencing data obtained through high-throughput sequencing technology,screen differential miRNAs with the standard of P<0.05,use miRanda and RNAhybrid to obtain differentially expressed miRNA candidate target genes,perform GO,KEGG enrichment analysis,and screen The biological processes,cell components,molecular functions and related signal pathways involved in it are listed.Results:(1)There were 52 differential miRNAs in the model group com pared to the normal control group,48 differential miRNAs that met the prediction requirements,and a Total of 1530 target genes were predict ed.The miRNAs with up-regulated expression are rno-miR-375-3p,rnomiR-292-3p,rno-miR-129-1-3p,rno-miR-129-2-3p,rno-miR-291a-3p,et c.The down-regulated miRNAs are rno-miR-144-3p,rno-miR-542-3p,rn o-miR-144-5p,etc.The biological processes involved mainly involve pept idyl-threonine phosphorylation modification and cellular metabolic proces ses Regulation,regulation of the process of dendritic cell apoptosis;cell components mainly include intracellular organelles,cytoplasm,actin cyt oskeleton,and cytoskeleton parts;molecular functions mainly involve tran sfeRase activity,small molecule binding,RNA polymeRase Ⅲ transcrip tion The sequence-specific DNA and other aspects of the regulatory reg ion.The KEGG signal pathways involved are Ras signal pathway,MAP K signal pathway,PI3K-Akt signal pathway,AMPK signal pathway.(2)There are a Total of 48 differential miRNAs in the traditional Chinese medicine group compared to the model group,46 differential miRNAs t hat meet the prediction requirements,and a total of 2124 target genes.S ignificant miRNAs represented by rno-miR-375-3p,rno-miR-292-3p,rno-miR-129-1-3p,rno-miR-129-2-3p,rno-miR-291 a-3p,etc.Up-regulation,miRNAs represented by rno-miR-144-3p,rno-miR-542-3p,rno-miR-1445p,etc.were significantly down-regulated.Biological processes mainly i nvolve the regulation of cell communication,signal transduction,progra mmed cell death,etc.;cell components mainly include organelles,cytopl asm,protein complexes,synapses,etc.;molecular functions mainly inclu de enzyme binding and RNA polymeRase Ⅲ regulatory regions DNA b inding,protein kinase binding,protein serine/threonine phosphatase inhi bitor activity,etc.The KEGG signaling pathways involved include AMP K signaling pathway,Ras signaling pathway,mTOR signaling pathway,NF-κB signaling pathway and so on.(3)There are 42 different genes in the western medicine group compared with the model group,including 20 up-regulated and 22 down-regulated,resulting in a Total of 1129 tar get genes.GO enrichment analysis mainly involves the organization of c ellular components,the organization of cellular components,or biogene sis,Protein location in the membrane,regulation of cell components an d tissues and other biological processes;cell components such as intracel lular parts,intracellular organelles,membrane-bound organelles,organel les,and intracellular membrane-bound organelles;enzyme binding,RNA polymeRase Ⅲ transcription regulation region Molecular functions such as sequence-specific DNA binding and transfeRase activity.KEGG enrich ment analysis mainly involves axon guidance,mTOR signaling pathway,AMPK signaling pathway and so on.Conclusion:(1)The incidence of thin endometrium may be related to rn o-miR-375-3p,rno-miR-292-3p,rno-miR-129-1-3p,rno-miR-129-2-3p,r no-miR-291a-3p,rno-miR-144-3p,rno-miR-542-3p,rno-miR-144-5p an d other miRNA expression related,the KEGG signaling pathways that may be involved include Ras signaling pathway,MAPK signaling path way,PI3K-Akt signal pathway,AMPK signal pathway,etc.(2)It is inferred that Bushen Tiaochong Decoction may regulate rno-miR-292-3p,rno-miR-292-5p,rno-miR-291 a-3p,rno-miR-291b,rno-let-7c-13p.The expression of miRNAs such as rno-miR-383-5p and rno-miR-338-5p acts on thin endometrial model rats,which may be involved in the regulation of AMPK signaling pathway,Ras signaling pathway,mTOR signaling pathway,and NF-κB signaling pathway.The pathway participat es in important processes such as endometrial proliferation of thin endo metrium,establishment of endometrial receptivity,and embryo implantat ion.(3)The effect of estradiol valerate on the thin endometrium may affect t he expression of miRNAs such as rno-miR-290-295 cluster,rno-let-7c-13p,miR-383-5p,and implantation from embryos.The aspects of estroge n production,oogenesis,inflammation,and development affect the prol iferation of the thin endometrium and the process of embryo implantati on.The KEGG signaling pathway,mTOR signaling pathway,AMPK sig naling pathway,etc.may be involved.