Differential expression of microRNAs in serum exosomes of obese and non-obese mice and analysis of their function

• Published in: Gene 2024-11, Vol.927, p.148604, Article 148604
• Main Authors: Wang, Changzan, Li, Xianghui, Yi, Wenying, Kang, Jiawei, Nuermaimaiti, Nuerbiye, Guan, Yaqun
• Format: Article
• Language: English
• Subjects: Animals; Disease Models, Animal; Exosomes - genetics; Exosomes - metabolism; Gene Expression Profiling - methods; Gene Expression Regulation; High-Throughput Nucleotide Sequencing; Insulin resistance; Male; Mice; Mice, Inbred C57BL; Mice, Obese; microRNA sequencing; MicroRNAs - blood; MicroRNAs - genetics; Obesity - blood; Obesity - genetics; Obesity - metabolism; Obesity exosome; Target point
• ISSN: 0378-1119; 1879-0038; 1879-0038
• DOI: 10.1016/j.gene.2024.148604

•In this study, serum exosomes from obese and non-obese mice were sequenced for the first time, and several differentially expressed microRNAs that may be involved in glycolipid metabolism were screened and validated.•This study discusses whether miR-674-5p can mediate the regulation of insulin resistance by IGF-II through the PI3K/Akt signalling pathway, pointing out the direction for subsequent studies.•In this study, GO analysis and KEGG analysis were performed to investigate the major enrichment sites of differentially expressed genes. To extract exosomes from obese and non-obese mice, screen specifically expressed microRNAs by high-throughput sequencing and explore their roles. An animal obesity model was constructed, and the successful construction of the obesity model was verified by HE staining, Western Blot and RT-qPCR. In addition, exosomes were extracted and verified by Western Blot. High-throughput sequencing was performed on the extracted serum exosomes to screen for differentially expressed microRNAs. fluorescence quantitative RT-PCR (RT-qPCR) was used to validate the differentially expressed miRNAs and explore their functions. 8 microRNAs were up-regulated and 11 microRNAs were down-regulated. mmu-miR-674-5p and X_28316 were significantly down-regulated and had the greatest impact on protein pathways. 8_13258 was significantly up-regulated and affected multiple protein pathways. GO enrichment analysis suggested that the differentially expressed microRNAs were mainly involved in the cleavage of microtubule activity, transferase activity/transferase pentameric acid. GO enrichment analysis suggested that differentially expressed microRNAs were mainly involved in the processes of cleavage microtubule activity, transferase activity/transfer pentamer, and threonine phosphatase/threonine kinase activity.KEGG pathway enrichment analysis showed that differentially expressed microRNAs were mainly involved in the processes of regulating the phosphorylation of TP53 activity, the G2/M DNA damage checkpoint, and the processing of the ends of DNA double-strand breaks. Protein interaction networks were enriched for Stat3, Fgr, Camk2b, Rac1, Asb6, and Ankfy1. Suggesting that they may be mediated by differential genes to participate in the process of insulin resistance. qRT-PCR results showed that the expression trend of mmu-miR-674-5p was consistent with the sequencing results. It suggests that it may be able to participate in the regulation of insulin resis