2,4-Thiazolidinedione in Well-Fed Lactating Dairy Goats: I. Effect on Adiposity and Milk Fat Synthesis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Design and Animal Management
2.2. Treatments
2.3. Measurements, Sample Collection, and Blood Metabolites
2.4. Adipose Biopsy and Histological Analysis, and Mammary Epithelial Cells Isolation
2.5. Reverse Transcriptase Quantitative Polymerase Chain Reaction (RT-qPCR)
2.6. Statistical Analysis
3. Results
3.1. Animal Performance
3.2. Blood Metabolic Parameters
3.3. Adipocytes Size Distribution
3.4. Gene Expression
3.4.1. Subcutaneous Adipose Tissue
3.4.2. Mammary Epithelial Cells
3.5. Fatty Acid Profile in Milk
4. Discussion
4.1. 2,4-TZD Induces Greater Glucose Utilization in the Adipose Tissue but Did Not Activate PPARγ
4.2. 2,4-TZD Does Not Affect Milk Fat Synthesis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A. Supplementary Materials and Methods
Appendix A.1. Animal Management
Appendix A.2. Dry Matter Intake, Energy Corrected Milk (ECM), Dairy Efficiency (DE), and Body Condition Score (BCS)
Appendix A.3. Adipose Biopsy
Appendix A.4. Histological Analysis of the Adipose Tissue
Appendix A.5. Mammary Epithelial Cells Isolation
Appendix A.6. RT-qPCR
Appendix A.7. Milking and Milk Sample Analysis
Appendix A.8. Milk Fatty Acid Analysis
Appendix A.9. Blood Metabolites
Appendix A.10. Statistical Analysis
References
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Parameter | Group | Day of Treatment | SEM | p-Value 1 | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
0 | 2 | 5 | 15 | 16 | 17 | 20 | 21 | 22 | Z | Time | Z × T | |||
Protein % | CTR | 2.32 | 2.29 | 2.22 | 2.32 | 2.35 | 2.28 | 2.41 | 2.41 | 2.39 | 0.09 | 0.30 | 0.10 | 0.78 |
TZD | 2.32 | 2.26 | 2.19 | 2.18 | 2.18 | 2.17 | 2.21 | 2.21 | 2.19 | 0.09 | ||||
Protein g 2 | CTR | 18.3 | 19.4 | 18.2 | 20.3 | 18.2 | 17.0 | 19.2 | 18.4 | 18.1 | 2.5 | 0.98 | <0.05 | 0.76 |
TZD | 18.3 | 20.4 | 17.9 | 18.9 | 18.2 | 17.5 | 18.4 | 19.2 | 19.0 | 2.5 | ||||
Lactose % | CTR | 4.45 | 4.47 | 4.42 | 4.38 | 4.40 | 4.35 | 4.34 | 4.35 | 4.31 | 0.07 | 0.45 | <0.05 | 0.23 |
TZD | 4.45 | 4.68 | 4.51 | 4.33 | 4.37 | 4.41 | 4.42 | 4.39 | 4.37 | 0.07 | ||||
Lactose g 2 | CTR | 34.9 | 37.2 | 36.0 | 37.8 | 33.7 | 32.1 | 34.2 | 32.7 | 32.2 | 4.3 | 0.61 | <0.05 | 0.61 |
TZD | 34.9 | 40.3 | 36.6 | 37.3 | 36.4 | 35.5 | 36.9 | 38.1 | 37.9 | 4.3 | ||||
SNF 3% | CTR | 7.66 | 7.61 | 7.55 | 7.53 | 7.51 | 7.45 | 7.48 | 7.51 | 7.46 | 0.31 | 0.98 | 0.44 | 0.63 |
TZD | 7.66 | 7.87 | 7.61 | 7.42 | 7.44 | 7.46 | 7.46 | 7.43 | 7.40 | 0.31 | ||||
SNF g 2 | CTR | 60.2 | 64.6 | 61.5 | 65.4 | 57.8 | 55.1 | 59.2 | 56.8 | 56.0 | 7.6 | 0.69 | <0.05 | 0.67 |
TZD | 60.2 | 69.3 | 62.0 | 64.1 | 62.1 | 60.2 | 62.1 | 64.5 | 64.2 | 7.6 |
Parameter | Group | Day of Treatment | SEM | p-Value 1 | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
0 | 7 | 9 | 15 | 16 | 17 | 18 | 21 | 26 | Z | Time | Z × T | |||
Cholesterol, mM | CTR | 2.55 | 2.30 | 2.27 | 2.18 | 2.28 | 2.31 | 2.33 | 2.40 | 2.32 | 0.24 | 0.38 | 0.18 | 0.70 |
TZD | 2.55 | 2.73 | 2.56 | 2.40 | 2.51 | 2.61 | 2.61 | 2.62 | 2.61 | 0.24 | ||||
∆FA, % 3 | CTR | 0.0 | 5.1 | −6.8 | 5.0 | 3.8 | 0.8 | 9.1 | 7.7 | −6.6 | 8.5 | 0.06 | 0.88 | 0.90 |
TZD | 0.0 | −13.2 | −14.5 | −9.4 | −4.9 | −8.1 | −9.7 | −17.2 | −12.0 | 8.5 | ||||
TAG, mM | CTR | 0.14 | 0.15 | 0.16 | 0.19 | 0.17 | 0.18 | 0.19 | 0.19 | 0.13 | 0.02 | 0.89 | 0.30 | 0.82 |
TZD | 0.14 | 0.14 | 0.15 | 0.19 | 0.19 | 0.19 | 0.18 | 0.16 | 0.15 | 0.02 | ||||
Creatinine, µM | CTR | 71.9 | 77.3 | 77.1 | 71.9 | 72.6 | 70.2 | 70.4 | 72.2 | 75.8 | 2.7 | 0.98 | <0.05 | 0.94 |
TZD | 71.9 | 76.6 | 77.6 | 70.7 | 72.2 | 71.7 | 70.6 | 72.4 | 76.1 | 2.7 | ||||
Urea, mM | CTR | 6.73 | 6.32 | 4.59 | 5.97 | 6.81 | 5.61 | 6.54 | 6.45 | 8.05 | 0.45 | 0.25 | <0.05 | 0.19 |
TZD | 6.73 | 6.40 | 5.16 | 5.29 | 6.08 | 5.08 | 5.61 | 5.71 | 6.22 | 0.45 |
Gene | Group | Time | SEM 1 | p-Value 2 | ||||
---|---|---|---|---|---|---|---|---|
−1 | 13 | 26 | Z | Time | Z × T | |||
Fatty Acids Import and De Novo Fatty Acid Synthesis | ||||||||
CD36 | CTR | 7169.8 | 6987.6 | 6233.3 | 681.3 | 0.03 | 0.03 | 0.03 |
TZD | 7169.8 | 3362.5 | 5781.9 | |||||
LPL | CTR | 122.2 | 130.5 | 244.6 | 52.1 | 0.62 | 0.04 | 0.90 |
TZD | 122.2 | 118.0 | 206.7 | |||||
SLC27A6 | CTR | 0.09 | 0.12 | 0.11 | 0.03 | 0.84 | 0.15 | 0.94 |
TZD | 0.09 | 0.14 | 0.11 | |||||
VLDLR | CTR | 2.3 | 2.0 | 1.0 | 0.9 | 0.67 | 0.48 | 0.25 |
TZD | 2.3 | 1.1 | 2.7 | |||||
ACACA | CTR | 37.6 | 26.6 | 48.5 | 24.3 | 0.33 | 0.51 | 0.14 |
TZD | 37.6 | 88.6 | 45.6 | |||||
FASN | CTR | 473.3 | 47.7 | 58.6 | 439.9 | 0.13 | 0.29 | 0.18 |
TZD | 473.3 | 163.1 | 1328.2 | |||||
SCD1 | CTR | 275.2 | 500.8 | 410.9 | 333.4 | 0.67 | 0.16 | 0.54 |
TZD | 275.2 | 917.0 | 329.3 | |||||
Insulin Signal and Glucose Metabolism | ||||||||
GAPDH | CTR | 159.2 | 125.6 | 91.2 | 41.7 | 0.01 | 0.25 | 0.08 |
TZD | 159.2 | 271.2 | 195.9 | |||||
INSR | CTR | 7.8 | 5.2 | 3.7 | 1.9 | 0.12 | 0.37 | 0.21 |
TZD | 7.8 | 5.8 | 9.4 | |||||
IRS1 | CTR | 9.2 | 6.8 | 4.6 | 2.0 | 0.04 | 0.42 | 0.05 |
TZD | 9.2 | 7.4 | 12.5 | |||||
SLC2A4 | CTR | 4.5 | 5.6 | 5.1 | 1.6 | 0.09 | 0.10 | 0.40 |
TZD | 4.5 | 8.9 | 7.3 | |||||
YWHAZ | CTR | 167.5 | 191.8 | 149.0 | 23.9 | 0.07 | 0.50 | 0.04 |
TZD | 167.5 | 101.4 | 150.1 | |||||
Glyceroneogenesis | ||||||||
PCK1 | CTR | 8.0 | 9.8 | 15.0 | 4.2 | 0.67 | 0.01 | 0.47 |
TZD | 8.0 | 8.7 | 21.0 | |||||
PDK4 | CTR | 1.8 | 1.2 | 1.1 | 0.4 | 0.66 | 0.03 | 0.08 |
TZD | 1.8 | 0.8 | 2.1 | |||||
GPD1 | CTR | 71.5 | 109.1 | 90.8 | 33.9 | 0.45 | 0.07 | 0.71 |
TZD | 71.5 | 145.8 | 116.4 | |||||
Transcription Regulation | ||||||||
PPARG | CTR | 43.5 | 41.7 | 49.5 | 9.6 | 0.81 | 0.19 | 0.70 |
TZD | 43.5 | 37.3 | 58.3 | |||||
SREBF1 | CTR | 16.1 | 19.3 | 23.0 | 16.7 | 0.61 | 0.38 | 0.61 |
TZD | 16.1 | 14.8 | 44.7 |
Gene | Group | Cell 1 | SEM | p-Value 2 | |||
---|---|---|---|---|---|---|---|
Neg | Pos | Z | Cell | Z × Cell | |||
Mammary Epithelial-Specific Genes | |||||||
CSN3 | CTR | 6324 | 3877 | 875 | 0.46 | <0.05 | 0.42 |
TZD | 5050 | 3326 | 911 | ||||
FABP3 | CTR | 20.0 | 8.5 | 3.0 | 0.57 | <0.05 | 0.41 |
TZD | 16.1 | 7.8 | 3.2 | ||||
LALBA | CTR | 1341 | 578 | 233.4 | 0.56 | <0.05 | 0.56 |
TZD | 1065 | 501 | 251.7 | ||||
KRT8 (Ln) 3 | CTR | 1.78 | 2.50 | 0.27 | 0.30 | <0.05 | 0.64 |
TZD | 1.48 | 2.03 | 0.29 | ||||
Fatty Acid Transport and Synthesis | |||||||
LPL | CTR | 9.68 | 4.87 | 1.52 | 0.70 | <0.05 | 0.77 |
TZD | 10.8 | 5.19 | 1.68 | ||||
CD36 | CTR | 1300 | 979 | 104 | 0.30 | <0.05 | 0.87 |
TZD | 1445 | 1094 | 115 | ||||
VLDLR (Ln) | CTR | 0.57 | 0.84 | 0.13 | 0.21 | <0.05 | 0.31 |
TZD | 0.43 | 0.53 | 0.14 | ||||
ACACA | CTR | 0.29 | 0.58 | 0.09 | 0.99 | 0.09 | 0.10 |
TZD | 0.43 | 0.43 | 0.10 | ||||
SCD1 | CTR | 5.84 | 5.03 | 0.68 | 0.06 | 0.02 | 0.53 |
TZD | 4.19 | 2.89 | 0.71 | ||||
Transcriptional Regulation | |||||||
NFE2L2 | CTR | 51.2 | 62.0 | 12.3 | 0.30 | <0.05 | 0.50 |
TZD | 66.4 | 83.7 | 12.6 | ||||
NRF1 | CTR | 0.15 | 0.17 | 0.03 | 0.69 | 0.50 | 0.91 |
TZD | 0.17 | 0.18 | 0.03 | ||||
PPARG (Ln) | CTR | -2.73 | −1.94 | 0.33 | 0.38 | <0.05 | 0.50 |
TZD | -2.20 | −1.71 | 0.35 | ||||
SREBF1 | CTR | 0.85 | 1.17 | 0.13 | 0.45 | <0.05 | 0.79 |
TZD | 0.74 | 1.01 | 0.14 | ||||
Other | |||||||
GAPDH (Ln) | CTR | 3.63 | 4.07 | 0.31 | 0.39 | <0.05 | 0.47 |
TZD | 3.85 | 4.56 | 0.33 | ||||
IL8 (Ln) | CTR | 4.8 | 4.9 | 0.6 | 0.31 | 0.17 | 0.40 |
TZD | 5.5 | 5.9 | 0.6 | ||||
YWHAZ | CTR | 31.6 | 40.7 | 8.3 | 0.64 | 0.19 | 0.61 |
TZD | 39.1 | 43.3 | 8.3 |
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Jaaf, S.; Rosa, F.; Moridi, M.; Osorio, J.S.; Lohakare, J.; Trevisi, E.; Filley, S.; Cherian, G.; Estill, C.T.; Bionaz, M. 2,4-Thiazolidinedione in Well-Fed Lactating Dairy Goats: I. Effect on Adiposity and Milk Fat Synthesis. Vet. Sci. 2019, 6, 45. https://doi.org/10.3390/vetsci6020045
Jaaf S, Rosa F, Moridi M, Osorio JS, Lohakare J, Trevisi E, Filley S, Cherian G, Estill CT, Bionaz M. 2,4-Thiazolidinedione in Well-Fed Lactating Dairy Goats: I. Effect on Adiposity and Milk Fat Synthesis. Veterinary Sciences. 2019; 6(2):45. https://doi.org/10.3390/vetsci6020045
Chicago/Turabian StyleJaaf, Shana, Fernanda Rosa, Misagh Moridi, Johan S. Osorio, Jayant Lohakare, Erminio Trevisi, Shelby Filley, Gita Cherian, Charles T. Estill, and Massimo Bionaz. 2019. "2,4-Thiazolidinedione in Well-Fed Lactating Dairy Goats: I. Effect on Adiposity and Milk Fat Synthesis" Veterinary Sciences 6, no. 2: 45. https://doi.org/10.3390/vetsci6020045