D-Limonene Promotes Anti-Obesity in 3T3-L1 Adipocytes and High-Calorie Diet-Induced Obese Rats by Activating the AMPK Signaling Pathway
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Cell Culture and Adipocyte Differentiation
2.3. Cell Viability Assay
2.4. Intracellular Lipid Assay
2.4.1. Oil Red O Staining Assay
2.4.2. Intracellular Triglyceride Assay
2.5. Animals and Diet
2.6. Animal Growth Characteristics Assay
2.7. Biochemical Parameters Assay
2.8. Histological Assay
2.9. mRNA Expression Assay
2.10. Protein Expression Assay
2.11. Statistical Assay
3. Results
3.1. Effect of LIM on Cell Viability and Proliferation Ability of 3T3-L1 Adipocytes
3.2. Effect of LIM on Lipid Accumulation in 3T3-L1 Adipocytes
3.3. Effect of LIM on Protein and mRNA Expression Related to AMPK Signaling Pathway in 3T3-L1 Adipocytes
3.4. Effect of LIM on the Growth Characteristics in High-Calorie Diet-Induced Obese Rats
3.5. Effect of LIM on Biochemical Parameters in High-Calorie Diet-Induced Obese Rats
3.6. Effect of LIM on Epididymal Adipose Tissue Sections in High-Calorie Diet-Induced Obese Rats
3.7. Effect of LIM on mRNA Expression of Genes Related to AMPK Signaling Pathway in High-Calorie Diet-Induced Obese Rats
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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ND | HD | HD + LLIM | HD + HLIM | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Body weight (g) | 493.50 | ± | 9.32 b | 564.25 | ± | 47.14 a | 503.13 | ± | 42.60 b | 477.23 | ± | 41.07 b |
Weight gain (%) | 94.15 | ± | 4.39 c | 123.38 | ± | 16.38 a | 111.01 | ± | 3.63 b | 100.37 | ± | 6.52 bc |
Lee’s index (g/cm) | 0.30 | ± | 0.01 c | 0.35 | ± | 0.01 a | 0.32 | ± | 0.00 b | 0.30 | ± | 0.00 c |
Food intake (g/rat/day) | 25.89 | ± | 1.07 a | 18.23 | ± | 0.96 b | 19.02 | ± | 1.11 b | 19.11 | ± | 0.87 b |
Feed conversion ratio (%) | 8.77 | ± | 0.37 d | 16.12 | ± | 1.21 a | 13.41 | ± | 0.36 b | 12.04 | ± | 0.57 c |
Liver (%) | 3.46 | ± | 0.30 a | 3.22 | ± | 0.27 a | 3.50 | ± | 0.41 a | 3.63 | ± | 0.44 a |
Kidney (%) | 0.81 | ± | 0.06 a | 0.65 | ± | 0.04 b | 0.69 | ± | 0.11 b | 0.82 | ± | 0.05 a |
Spleen (%) | 0.19 | ± | 0.01 a | 0.17 | ± | 0.02 a | 0.17 | ± | 0.01 a | 0.16 | ± | 0.02 a |
Epididymal adipose tissue (%) | 1.55 | ± | 0.18 b | 2.83 | ± | 0.94 a | 2.22 | ± | 0.46 ab | 1.83 | ± | 0.35 b |
Mesenteric adipose tissue (%) | 1.02 | ± | 0.04 c | 1.90 | ± | 0.15 a | 1.43 | ± | 0.20 b | 1.17 | ± | 0.16 c |
Retroperitoneal adipose tissue (%) | 1.23 | ± | 0.05 c | 3.16 | ± | 0.64 a | 2.04 | ± | 0.43 b | 2.05 | ± | 0.17 b |
Total fat tissue (%) | 3.81 | ± | 0.59 c | 8.25 | ± | 1.11 a | 5.78 | ± | 1.02 b | 5.24 | ± | 0.58 b |
ND | HD | HD + LLIM | HD + HLIM | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Triglyceride (mg/dL) | 75.11 | ± | 7.26 a | 84.05 | ± | 4.54 a | 74.10 | ± | 9.51 a | 45.79 | ± | 1.81 b |
Total cholesterol (mg/dL) | 78.84 | ± | 2.71 a | 72.36 | ± | 2.16 b | 77.95 | ± | 2.78 a | 73.29 | ± | 2.84 b |
Low-density lipoprotein cholesterol (mg/dL) | 9.50 | ± | 0.58 c | 12.67 | ± | 0.58 a | 11.00 | ± | 0.00 b | 11.67 | ± | 0.58 b |
High-density lipoprotein cholesterol (mg/dL) | 48.00 | ± | 5.48 ab | 41.67 | ± | 2.31 b | 43.67 | ± | 2.08 ab | 51.00 | ± | 6.00 a |
Free fatty acid (mmol/L) | 1.44 | ± | 0.12 a | 1.32 | ± | 0.23 a | 1.43 | ± | 0.06 a | 1.38 | ± | 0.13 a |
Lipase (mmol/L) | 7.20 | ± | 0.45 a | 6.40 | ± | 0.55 b | 6.57 | ± | 0.53 ab | 7.20 | ± | 0.45 a |
Ketone body (mmol/L) | 1.24 | ± | 0.21 a | 1.52 | ± | 0.13 a | 1.34 | ± | 0.15 a | 1.42 | ± | 0.16 a |
Aspartate aminotransferase (U/L) | 136.20 | ± | 28.60 a | 134.60 | ± | 7.64 a | 131.29 | ± | 12.40 a | 135.35 | ± | 27.32 a |
Alanine aminotransferase (U/L) | 50.20 | ± | 5.26 a | 49.40 | ± | 3.83 a | 45.79 | ± | 7.38 a | 43.67 | ± | 5.13 a |
Creatinine (mg/dL) | 0.71 | ± | 0.03 a | 0.75 | ± | 0.08 a | 0.67 | ± | 0.06 ab | 0.62 | ± | 0.06 b |
Uric acid (mg/dL) | 2.18 | ± | 0.33 b | 3.10 | ± | 0.56 a | 3.92 | ± | 0.59 a | 3.13 | ± | 0.57 a |
Sodium (mmol/L) | 150.82 | ± | 2.50 a | 150.33 | ± | 2.52 a | 150.69 | ± | 2.20 a | 149.65 | ± | 2.56 a |
Potassium (mmol/L) | 7.22 | ± | 1.61 a | 7.16 | ± | 0.67 a | 7.99 | ± | 0.79 a | 7.78 | ± | 0.80 a |
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Liao, J.-T.; Huang, Y.-W.; Hou, C.-Y.; Wang, J.-J.; Wu, C.-C.; Hsieh, S.-L. D-Limonene Promotes Anti-Obesity in 3T3-L1 Adipocytes and High-Calorie Diet-Induced Obese Rats by Activating the AMPK Signaling Pathway. Nutrients 2023, 15, 267. https://doi.org/10.3390/nu15020267
Liao J-T, Huang Y-W, Hou C-Y, Wang J-J, Wu C-C, Hsieh S-L. D-Limonene Promotes Anti-Obesity in 3T3-L1 Adipocytes and High-Calorie Diet-Induced Obese Rats by Activating the AMPK Signaling Pathway. Nutrients. 2023; 15(2):267. https://doi.org/10.3390/nu15020267
Chicago/Turabian StyleLiao, Jin-Ting, Yu-Wen Huang, Chih-Yao Hou, Jyh-Jye Wang, Chih-Chung Wu, and Shu-Ling Hsieh. 2023. "D-Limonene Promotes Anti-Obesity in 3T3-L1 Adipocytes and High-Calorie Diet-Induced Obese Rats by Activating the AMPK Signaling Pathway" Nutrients 15, no. 2: 267. https://doi.org/10.3390/nu15020267