Bta-miR-365-3p-targeted FKBP5 participates in the AMPK/mTOR signaling pathway in the regulation of preadipocyte differentiation in cattle

Mengdi Chen; Congcong Zhang; Zewen Wu; Siwei Guo; Wenfa Lv; Jixuan Song; Beibei Hao; Jinhui Bai; Xinxin Zhang; Hongyan Xu; Guangjun Xia

doi:10.5713/ab.23.0328

Bta-miR-365-3p-targeted FKBP5 participates in the AMPK/mTOR signaling pathway in the regulation of preadipocyte differentiation in cattle

Anim Biosci. 2024 Apr 24. doi: 10.5713/ab.23.0328. Online ahead of print.

Authors

Mengdi Chen^{1

2}, Congcong Zhang³, Zewen Wu¹, Siwei Guo¹, Wenfa Lv⁴, Jixuan Song¹, Beibei Hao^{1

2}, Jinhui Bai¹, Xinxin Zhang^{1

2}, Hongyan Xu^{1

2}, Guangjun Xia^{1

2

5}

Affiliations

¹ College of Agriculture, Yanbian University, Yanji 133000, China.
² College of Integration Science , Yanbian University, Yanji 133000,China.
³ Institute of Animal Science, Jilin Academy of Agricultural Science, Changchun 136100, China.
⁴ Joint Laboratory of Modern Agricultural Technology International Cooperation, Ministry of Education, Jilin Agricultural University, Changchun 130118, China.
⁵ Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Yanbian University, Yanji, Jilin, China.

PMID: 38665092
DOI: 10.5713/ab.23.0328

Abstract

Objective: MicroRNAs (miRNAs) are endogenous non-coding RNAs that can play a role in the post-transcriptional regulation of mammalian preadipocyte differentiation. However, the precise functional mechanism of its regulation of fat metabolism is not fully understood.

Methods: We identified bta-miR-365-3p, which specifically targets the 3'untranslated region (3'UTR) of the FK506-binding protein 5 (FKBP5),and verified its mechanisms for regulating expression and involvement in adipogenesis.

Results: In this study, we found that the overexpression of bta-miR-365-3p significantly decreased the lipid accumulation and triglyceride content in the adipocytes. Compared to inhibiting bta-miR-365-3p group, overexpression of bta-miR-365-3p can inhibit the expression of adipocyte differentiation-related genes C/EBPα and PPARγ. The dual-luciferase reporter system further validated the targeting relationship between bta-miR-365-3p and FKBP5. FKBP5 mRNA and protein expression were detected by qRT-PCR and Western blot. Overexpression of bta-miR-365-3p significantly down-regulated FKBP5 expression, while inhibition of bta-miR-365-3p showed the opposite, indicating that bta-miR-365-3p negatively regulates FKBP5. AMPK/mTOR signaling pathway is closely related to the regulation of cell growth and is involved in the development of bovine adipocytes. In this study, overexpression of bta-miR-365-3p significantly inhibited mRNA and protein expression of AMPK, mTOR, and SREBP1 genes, while the inhibition of bta-miR-365-3p expression was contrary to these results. Overexpression of FKBP5 significantly upregulated AMPK, mTOR, and SREBP1 gene expression, while inhibition of FKBP5 expression was contrary to the above experimental results.

Conclusion: In conclusion, these results indicate that bta-miR-365-3p may be involved in the AMPK/mTOR signaling pathway in regulating Yanbian yellow cattle preadipocytes differentiation by targeting the FKBP5 gene.

Keywords: AMPK/mTOR Signalling Pathway; FKBP5; Preadipocytes; Yanbian Yellow Cattle; bta-miR-365-3p.