Analysis Of Differential Expression Of Non-coding Small RNA During Browning Of White Adipose Tissue

Part1 Analysis of miRNAs’ differential expression during browning of white adipose tissue induced by different methods and its potential regulatory mechanismObjective To investigate the differential expressed miRNAs in the white adipose tissue browning of mice under different stimulation conditions,and then analyze their potential regulatory mechanisms.Method White adipose tissue browning mouse models were established in different groups,cold-induced group and injected CL316,243 group.HE staining and gene expression analysis of inguinal subcutaneous white adipose tissue(sWAT)were used for identifying the browning models.miRNAs expression profile in different conditions were described byRNA-sequencing(RNA-seq),miRNAs that have same expression trend in two groups were screened out.Target genes of miRNAs were predicted by bioinformatics,and their expression level were verified by quantitative real-time PCR(qRT-PCR).Results Both cold-induced and CL316,243 Intraperitoneal injection could activate the browning of sWAT,and miRNA expression profile of sWAT was significantly different after induction.The expression of miR-30e-3p was increased while the miR-181a-5p expression was reduced in the browning of sWAT.Prediction and validation of target genes revealed that Cdk6 and SIRT1 are potential targets regulated by miR-30e-3p and miR-181a-5p respectively in the browning of sWAT.Conclusions The miRNA expression difference of sWAT from the mice which are treatment cold stimulation and CL316,243 is significant.Mi R-30e-3p and miR-181a-5p may be involved in the regulation of sWAT browning process through target genes Cdk6 and Sirt1,respectively.Part2 Analysis of tsRNA expression changes during browning of white adipose tissueObjective To explore the differential expression of tsRNAs in the white adipose tissue of mice under different stimulation conditions and to analyze their potential molecular regulatory mechanism during browning.Method To build a brown model of white adipose tissue subcutaneously in mice,one group was hypothermic and the other group was intraperitoneal injection of CL316,243.Through HE staining and gene expression analysis related to thermogenesis,the establishment of a browning model of subcutaneous white adipose tissue was determined.RNA sequencing was used to obtain tsRNAs expression profiles of subcutaneous white adipose tissue under different conditions,and tsRNAs with the same differential expression trend in the two groups were screened and verified by real-time fluorescence quantitative PCR;tsRNAs target genes were predicted by bioinformatics and real-time fluorescence quantification PCR further verified its expression level.Results Both cold stimulation and CL316,243 can induce browning of subcutaneous white adipose tissue.At the same time,the expression profile of tsRNAs in subcutaneous white adipose tissue is significantly different before and after induction.The length distribution of tsRNAs and the type of precursor-derived tRNA are closely related toWAT browning.Enrichment analysis of tsRNAs target genes showed that tsRNA may regulateWAT browning through PI3K-Akt pathways.Conclusions The expression profile of tsRNAs in subcutaneous white adipose tissue was significantly different after browning induction.The results of target gene enrichment analysis showed that tsRNA may regulateWAT browning through PI3K-Akt and other pathways.