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FOXO3a regulates lipid accumulation and adipocyte inflammation in adipocytes through autophagy

Role of FOXO3a in obesity

  • Original Research Paper
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Abstract

Background

FOXO3a is a widely studied transcription factor and plays an important role in a variety of biology. The purpose of this study was to explore the role and potential mechanism of FOXO3a on lipid accumulation and adipocyte inflammation in adipocytes through regulation of autophagy.

Methods

The obese mouse model was successfully induced by high-fat diet. SiRNA targeting FOXO3a was transfected into differentiation of 3T3-L1 adipocytes to reduce the expression of FOXO3a. The culture medium of RAW264.7 cells was added to the differentiated 3T3-L1 adipocytes to form a co-culture system. Subsequently, ELISA or AdipoRed assay was performed to measure the expression of triglyceride (TG) and cholesterol (TC) in mouse adipose tissue or differentiation of 3T3-L1 adipocytes. Adipocyte differentiation was detected by Oil Red O-staining. Ad-mCherry-GFP-LC3II was used to detect the level of autophagy in differentiation of 3T3-L1 adipocytes. Western blotting or qRT-PCR was used to detect the expression of FOXO3a, autophagy-related proteins (beclin 1, CEBPβ, PPARγ, ACC1 and KLF4), inflammatory cytokines (TNF-α, IL-1β, IL-6 and MCP1), NF-κB signal pathway-related proteins or adipokines (Adiponectin, AdipoR1 and resistin) in differentiated 3T3-L1 or RAW264.7 cells.

Results

The expression of FOXO3a and autophagy levels were significantly increased in visceral adipose tissue of obese mice and differentiation of 3T3-L1 adipocytes. Downregulation of FOXO3a significantly inhibited the autophagy and lipid accumulation in differentiation of 3T3-L1 adipocytes. In addition, FOXO3a knockdown significantly reduced Lipopolysaccharide (LPS)-induced inflammation and adipokines release in RAW264.7 cells treated with the culture medium of 3T3-L1 adipocytes. These above activity changes could be reversed by autophagy inducer rapamycin.

Conclusion

FOXO3a could promote lipid accumulation and inflammation in differentiated 3T3-L1 adipocytes by targeting autophagy. Our results provide a new theoretical basis for FOXO3a to regulate obesity.

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The experimental data will be available on the request.

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Funding

Natural Science Foundation of Shanghai (18ZR1435800).

Author information

Author notes

  1. Xiaoyan Zhang and Qiang Liu contributed equally to this work.

Authors and Affiliations

  1. Department of Endocrinology, Yangpu Hospital, Tongji University School of Medicine, 450 Tengyue Road, Yangpu, Shanghai, 200090, China

    Xiaoyan Zhang, Xuane Zhang, Kai Guo, Xuelian Zhang & Zunhai Zhou

  2. Department of Cardiac Surgery, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, 200072, China

    Qiang Liu

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  1. Xiaoyan Zhang
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  2. Qiang Liu
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  3. Xuane Zhang
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  4. Kai Guo
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  5. Xuelian Zhang
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  6. Zunhai Zhou
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Contributions

Xiaoyan Zhang: project development, data analysis and collection, manuscript writing. QL: project development, data analysis and collection, manuscript writing. Xuane Zhang: project development, data collection. KG: project development, data collection. Xuelian Zhang: project development, manuscript editing. ZZ: project development, manuscript editing.

Corresponding author

Correspondence to Zunhai Zhou.

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The authors declare they have no competing interests.

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All the above experimental operations have been approved by the ethics committee of the Yangpu Hospital, Tongji University School of Medicine.

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Responsible Editor: John Di Battista.

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Zhang, X., Liu, Q., Zhang, X. et al. FOXO3a regulates lipid accumulation and adipocyte inflammation in adipocytes through autophagy. Inflamm. Res. 70, 591–603 (2021). https://doi.org/10.1007/s00011-021-01463-0

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  • DOI: https://doi.org/10.1007/s00011-021-01463-0

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