miR-191-5p Targets C/EBPβ to Inhibit Lipid Deposition in C2C12 Cells
LIAN Xinrong1,2, CHEN Haimin1, WANG Jiarui2, XIA Jingwen2, ZHANG Jin2,3*, WU Wenjing2*
The aim of this study is to investigate the effects and mechanisms of miR-191-5p on myogenic differentiation and ectopic adipogenesis in C2C12 cells. Useing mouse myoblast cell line C2C12 as model, the dif ferentiate into myotubes or to undergo intramyocellular lipid accumulation using horse serum, sodium oleate, and sodium palmitate were conducted. After transfection with miR-191-5p mimic or inhibitors, qRT-PCR and Western blot were performed to assess the expression of myogenic markers (MyoD, MyoG, MyHC, Myf5) and adipogenic markers (FAS, PPARγ, HSL), as well as to evaluate the effects on relevant signaling pathways. Furthermore, bio informatics analysis and dual-luciferase reporter assays were used to predict and validate the target genes of miR 191-5p. The results showed that miR-191-5p was highly expressed in mouse skeletal muscle and exhibited an initial increase followed by a decrease during C2C12 myogenic differentiation, suggesting its involvement in the regula tion of skeletal muscle development. Compared to the control group, overexpression of miR-191-5p promoted myogenic differentiation in C2C12 cells, significantly increasing the expression of MyoD, MyoG, and other myogenic genes. Regarding ectopic adipogenesis, overexpression of miR-191-5p significantly inhibited lipid droplet accumu lation and downregulated the expression of FAS and PPARγ. Additionally, phosphorylation of ERK1/2 was mark edly reduced. Co-transfection with psi-CHECK-2-C/EBPβ 3ʹUTR and miR-191-5p mimic resulted in a significant decrease in the ratio of firefly to Renilla luciferase activity, indicating that miR-191-5p directly targets the 3ʹUTR of C/EBPβ. In conclusion, miR-191-5p may promote myogenesis and suppress ectopic lipid deposition in C2C12 cells by targeting C/EBPβ and inhibiting the ERK1/2 signaling pathway.
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