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RNA-Seq analyses in bta-miR-484 transfected adipocyte

ABSTRACT: To elucidate the role of bta-miR-484 in adipocytes. we compared expression profiles of mRNAs in control and bta-miR-484 overexpressed adipocytes by RNA se-quencing. A total of 51 up-regulated genes and 77 down-regulated genes were detected in adipocytes overexpressing bta-miR-484. GO annotation showed that differ-entially expressed genes (DEGs） enrichment positive regulation of glycogen biosynthetic process, positive regulation of glucose import and other biological processes. COG classification found that DEGs are rich in Amino acid transport and metabolism, Posttranslational modification, protein turnover, chaperones, Secondary metabolites bio-synthesis, transport and catabolism, General function prediction only, Signal transduc-tion mechanisms, and other items. KEGG enrichment analysis showed that DEGs were enriched in apoptosis, AMPK signaling pathway, calcium signaling pathway, Ras signaling pathway, MAPK signaling pathway, Wnt signaling pathway, PI3K-Akt signaling pathway, adipocyto-kine signaling pathway, insulin secretion , PPAR signaling pathway, etc.

ORGANISM(S): Bos taurus

PROVIDER: GSE238109 | GEO | 2023/07/29

REPOSITORIES: GEO

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Project description:Liver fibrosis is a common pathological process of various chronic liver diseases that can develop into liver cancer. MicroRNAs (miRNAs) are a king of non-coding RNA which are closely related to liver diseases. Thus, this research hope to explore the effect of miR-484 on liver fibrosis and reveal its mechanism. The miRNAs profiles were screened by microRNA sequencing and the location of miR-484 was identified by fluorescence in situ hybridization (FISH) in human liver fibrotic tissues. MiR-484 expression was detected by qRT-PCR in rat primary hepatic stellate cells (HSCs). Bioinformatics analysis and dual-luciferase reporter assay were performed to determine the target gene of miR-484. Liver fibrosis specific signatures were analyzed by qRT-PCR and western blot after miR-484 mimic/inhibitor transfection. The cell apoptosis was detected by Annexin V-FITC/PI double staining. The effect of miR-484 silencing on fibrosis in vivo was investigated in thioacetamide (TAA) induced mice model using the adeno-associated virus carrying miR-484 inhibitor. Enrichment of miR-484 was observed in human liver fibrosis tissues and activated rat primary HSCs. FISH showed that miR-484 was prominently located at fibrotic region and the cytoplasm of HSCs in human liver tissues. Dual-luciferase reporter assay verified that the homeodomain-interacting protein kinases 1 (HIPK1) was the direct target of miR-484. After transfecting miR-484 inhibitor into HSC-T6, HIPK1 were significantly up-regulated, and α-SMA, col1a1, Wnt-3a, Wnt-5a, β-catenin and p-β-catenin were down-regulated, suggesting the restrain effect of miR-484 knockdown on HSCs activation. Conversely, the results were opposite with miR-484 mimic transfection. In addition, the apoptosis of HSC-T6 altered significantly after miR-484 modulation. Moreover, adeno-associated virus carrying miR-484 inhibitor alleviated mice liver fibrosis induced by TAA. In conclusion, miR-484 knockdown ameliorates liver fibrosis by promoting the apoptosis and suppressing HSCs activation via blocking Wnt/β-catenin signaling pathway. MiR-484 and its downstream gene HIPK1 might be selected as novel therapeutic targets of liver fibrosis.

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RNA-Seq analyses in bta-miR-484 transfected adipocyte

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