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Updated: Jun 19, 2026
Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice
Published on: November 27, 2019
Modelling 16/8 intermittent fasting and breakfast-skipping in mouse adipocytes 3T3-L1 in vitro: a transcriptomics
Pei Han Er1, Jin Yi Chye1, Geetha Letchumanan1
1Department of Biomedical Sciences, Sir Jeffrey Cheah Sunway Medical School, Faculty of Medical and Life Sciences, Sunway University, 5 Jalan Universiti, Bandar Sunway, 47500 Subang Jaya, Selangor, Malaysia.
Background:
Obesity, a chronic metabolic disease, lacks effective treatments. Intermittent fasting (IF), especially time-restricted feeding (TRF), shows promise, yet the comparative molecular effects of different regimens remain unclear. This study examined their impact on 3T3-L1 adipocytes and transcriptomic changes.
Methods:
3T3-L1 cells were differentiated into adipocytes for 7 d and synchronized with 200 nM dexamethasone before 24 h treatments: (1) Control [high glucose (4.5 g/L DMEM), 15 % bovine calf serum (BCS)], (2) 16 h fasting/8h feeding IF [control medium ZT3-ZT11; low glucose (1.0 g/L) and low serum (1 % BCS) ZT12-ZT2], (3) "Distributed" IF [medium glucose (2.75 g/L), medium serum (8 % BCS) ZT0-ZT24], (4) Breakfast-skipping (BS) [low glucose/low serum ZT1-ZT5; control medium ZT6-ZT0]. Lipid accumulation was assessed by Oil Red O staining; whole transcriptome sequencing was performed.
Results:
The 16/8 IF regimen produced the greatest lipid reduction (74.41 % vs. control; p = 0.007) with upregulation of lipolysis genes (Tgm2, Notch2) and downregulation of adipogenesis and glycolysis genes (Ccnd1, Ldha). Pathway enrichment included TGF-β, p53, and apelin signaling. In contrast, the BS regimen did not differ significantly in lipid accumulation compared to control (p = 0.999), but nevertheless showed downregulation of mitochondrial genes mt-Rnr1 and mt-Rnr2.
Conclusion:
This work highlights the novel comparative evaluation of IF/TRF regimens in adipocytes using transcriptomics, demonstrating that 16/8 IF most effectively reduced lipid content and modulated metabolic pathways within 24 h. These findings provide mechanistic insight that strengthens the rationale for TRF as a dietary approach for obesity management.
Insights
Intermittent fasting (IF), particularly 16/8 time-restricted feeding (TRF), significantly reduced adipocyte lipid content within 24 hours. This study provides molecular insights into TRF
Area of Science:
- Metabolic disease research
- Cell biology
- Molecular biology
Background:
- Obesity is a complex metabolic disease with limited effective treatments.
- Intermittent fasting (IF), especially time-restricted feeding (TRF), is a promising dietary strategy.
- Comparative molecular effects of different IF/TRF regimens require further investigation.
Purpose of the Study:
- To comparatively evaluate the molecular effects of different IF/TRF regimens on 3T3-L1 adipocytes.
- To identify transcriptomic changes induced by various IF/TRF protocols.
- To assess the impact of IF/TRF on lipid accumulation in adipocytes.
Main Methods:
- 3T3-L1 cells were differentiated into adipocytes and subjected to 24-hour treatments: control, 16/8 IF, 'distributed' IF, and breakfast-skipping (BS).
- Lipid accumulation was quantified using Oil Red O staining.
- Whole transcriptome sequencing was performed to analyze gene expression profiles.
Main Results:
- The 16/8 IF regimen demonstrated the most significant reduction in lipid accumulation (74.41%) compared to control.
- 16/8 IF upregulated lipolysis genes (e.g., Tgm2, Notch2) and downregulated adipogenesis/glycolysis genes (e.g., Ccnd1, Ldha).
- Breakfast-skipping showed no significant difference in lipid accumulation but downregulated mitochondrial genes.
Conclusions:
- The 16/8 IF regimen was most effective in reducing adipocyte lipid content and modulating metabolic pathways within 24 hours.
- Transcriptomic analysis revealed distinct molecular responses to different IF/TRF regimens.
- These findings support the rationale for TRF as a dietary intervention for obesity management.
