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Bta-miR-365-3p-targeted FK506-binding protein 5 participates in the AMPK/mTOR signaling pathway in the regulation of preadipocyte differentiation in cattle |
Mengdi Chen1,2,a 
, Congcong Zhang3,a
, Zewen Wu1
, Siwei Guo1
, Wenfa Lv4
, Jixuan Song1
, Beibei Hao1,2
, Jinhui Bai1
, Xinxin Zhang1,2
, Hongyan Xu1,2,*
, Guangjun Xia1,2,5,*
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1College of Agriculture, Yanbian University, Yanji 133000, China
2College of Integration Science, Yanbian University, Yanji 133000, China
3Institute of Animal Science, Jilin Academy of Agricultural Science, Changchun 136100, China
4Joint Laboratory of Modern Agricultural Technology International Cooperation, Ministry of Education, Jilin Agricultural University, Changchun 130118, China
5Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Yanbian University, Yanji, Jilin 133000, China |
Correspondence:
Hongyan Xu, Tel: +86-13944709169, Email: xuhongyan@ybu.edu.cn Guangjun Xia
Guangjun Xia, Tel: +86-13844705548, Email: ybuac@ybu.edu.cn
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Received: 29 August 2023 • Revised: 26 October 2023 • Accepted: 18 February 2024
aThese authors contributed equally to this work. |
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| 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-36 5-3p group, overexpression of bta-miR-365-3p can inhibit the expression of adipocyte differentiation-related genes C/EBPα and PPARγ. The dualluciferase reporter system further validated the targeting relationship between bta-miR-365-3p and FKBP5. FKBP5 mRNA and protein expression were detected by quantitative real-time polymerase chain reaction 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. Adenosine 5′-monophosphate (AMP)-activated protein kinase/mammalian target of rapamycin (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; bta-miR-365-3p; FK506-binding protein 5 (FKBP5); Preadipocytes; Yanbian Yellow Cattle |
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