Proteomics Analysis Reveals an Important Role for the PPAR Signaling Pathway in DBDCT-Induced Hepatotoxicity Mechanisms
Abstract
:1. Introduction
2. Results
2.1. Histopathology
2.2. Differentially Expressed Proteins in Rat Livers Exposed to DBDCT
2.3. Pathway Data
2.4. Cytotoxicities of DBDCT
2.5. DBDCT-Induced Apoptosis Mediated by Caspase Activation
2.6. DBDCT Regulated the mRNA Levels of Downstream Targets in the PPAR Signaling Pathway
2.7. Effects of DBDCT on Expressions of PPAR-α and PPAR-λ in Protein and Gene Levels
2.8. Effects of DBDCT on Expressions of PPAR-λ, CD36, FABP4 and EHHADH in Gene Levels
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Animals’ Treatment and Tissue Collection
4.3. Histological Evaluation of Liver Tissues
4.4. Sample Fractionation
4.5. Liquid Chromatography—Mass Spectrometry Analysis
4.6. Label-Free Protein Expression Data Processing
4.7. Cell Culture
4.8. Cytotoxicity Assay
4.9. Western Blot Analysis
4.10. RT-PCR Analysis
4.11. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are not available from the authors. |
Term | Count | p-Value |
---|---|---|
Oxidation reduction | 31 | 1.2 × 10−13 |
Response to organic substance | 21 | 6.9 × 10−4 |
Fatty acid metabolic process | 11 | 1.6 × 10−5 |
Lipid biosynthetic process | 10 | 1.8 × 10−3 |
Macromolecule catabolic process | 10 | 3.8 × 10−2 |
Response to hormone stimulus | 10 | 6.4 × 10−2 |
Protein transport | 10 | 9.2 × 10−2 |
Establishment of protein localization | 10 | 9.6 × 10−2 |
Response to drug | 9 | 1.5 × 10−2 |
Cofactor metabolic process | 8 | 3.8 × 10−3 |
Response to inorganic substance | 8 | 1.7 × 10−2 |
Protein catabolic process | 8 | 4.6 × 10−2 |
Lipid catabolic process | 7 | 2.1 × 10−3 |
Di- and tri-valent inorganic cation transport | 7 | 3.3 × 10−3 |
Steroid metabolic process | 7 | 6.3 × 10−3 |
Cellular protein catabolic process | 7 | 8.9 × 10−2 |
Organic acid biosynthetic process | 6 | 1.6 × 10−2 |
Carboxylic acid biosynthetic process | 6 | 1.6 × 10−2 |
Response to metal ion | 6 | 3.1 × 10−2 |
Response to organic cyclic substance | 6 | 5.7 × 10−2 |
Pathway | Count | p-Value |
---|---|---|
Drug metabolism | 13 | 2.8 × 10−11 |
Metabolism of xenobiotics by cytochrome P450 | 11 | 1.4 × 10−9 |
Retinol metabolism | 8 | 5.4 × 10−6 |
Linoleic acid metabolism | 6 | 3.9 × 10−5 |
PPAR signaling pathway | 7 | 1.8 × 10−4 |
Arachidonic acid metabolism | 6 | 1.4 × 10−3 |
Steroid biosynthesis | 4 | 1.7 × 10−2 |
β-Alanine metabolism | 3 | 2.8 × 10−2 |
Pyruvate metabolism | 3 | 7.9 × 10−2 |
Pyrimidine metabolism | 4 | 9.7 × 10−2 |
Protein ID | Protein Name | Fold Change | Protein Description |
---|---|---|---|
Q07969 | Platelet glycoprotein 4/CD36 | 3.56 | Binds long-chain fatty acids and may function in transport and/or as a regulator of fatty acid transport. As a coreceptor fortoll-like receptor 4-6 (TLR4-TLR6), promotes inflammation in monocytes/macrophages. Upon ligand binding, such as oxLDL or amyloid-β 42 binding, rapidly induces the formation of a heterodimer of TLR4 and TLR6, which is internalized and triggers inflammatory signals, leading to the NF-kappa-B-dependent production of CXCL1, CXCL2 and CCL9 cytokines via the MYD88 signaling pathway, and CCL5 cytokine via the TICAM1 signaling pathway, as well as IL1B secretion. |
P70623 | Fatty acid binding protein, adipocyte 4 | 2.02 | Lipid transport protein in adipocytes. Binds both long-chain fatty acids and retinoic acid. Delivers long-chain fatty acids and retinoic acid to their cognate receptors in the nucleus. |
P07896 | Peroxisomal bifunctional enzyme/enoyl-CoA hydratase | 3.16 | |
P07871 | 3-ketoacyl-CoA thiolase B, peroxisomal/acetyl-CoA acyltransferase 1 | 2.28 | |
P07379 | Phosphoenolpyruvate carboxykinase, cytosolic | 2.02 | Catalyzes the conversion of oxaloacetate (OAA) to phosphoenolpyruvate (PEP), the rate-limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle. |
Isoforms | Primer Sequence | Product Size (bp) | PCR Conditions | ||
---|---|---|---|---|---|
Cycle | Denaturation | Annealing | |||
β-actin | F:CTACAATGAGCTGCGTGTGGC R:CAGGTCCAGACGCAGGATGGC | 270 | 30 | 94 °C, 30 s | 55 °C, 30 s |
PPAR-α | F:ACGATGCTGTCCTCCTTGATG R:GCGTCTGACTCGGTCTTCTTG | 407 | 30 | 94 °C, 30 s | 52 °C, 30 s |
PPAR-λ | F:CCCTTTACCACGGTTGATTTCTC R:GCAGGCTCTACTTTGATCGCACD | 143 | 40 | 94 °C, 30 s | 59 °C, 30 s |
CD36 | F:TGAATCCTAACGAAGATGAGCA R:GGCTTGACCAGTATGTTGACC | 106 | 40 | 94 °C, 30 s | 60 °C, 30 s |
FABP4 | F:AGAAGTGGGAGTTGGCTTCG R:ACTCTCTGACCGGATGACGA | 103 | 40 | 94 °C, 30 s | 60 °C, 30 s |
ACAA1 | F:TGGCATCAGAAATGGGTCTT R:AGGAATCAGGCAGTCTCTGG | 136 | 40 | 94 °C, 30 s | 60 °C, 30 s |
EHHADH | F:CACGGTTATGAGCTTGTCCA R:TCTGGCTTGCTACCTTCCTC | 138 | 40 | 94 °C, 30 s | 60 °C, 30 s |
PEPCK | F:CTGCCTCTCTCCACACCATT R:GCCTTCCACAAACTTCCTCA | 125 | 40 | 94 °C, 30 s | 60 °C, 30 s |
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Li, Y.; Liu, X.; Niu, L.; Li, Q. Proteomics Analysis Reveals an Important Role for the PPAR Signaling Pathway in DBDCT-Induced Hepatotoxicity Mechanisms. Molecules 2017, 22, 1113. https://doi.org/10.3390/molecules22071113
Li Y, Liu X, Niu L, Li Q. Proteomics Analysis Reveals an Important Role for the PPAR Signaling Pathway in DBDCT-Induced Hepatotoxicity Mechanisms. Molecules. 2017; 22(7):1113. https://doi.org/10.3390/molecules22071113
Chicago/Turabian StyleLi, Yunlan, Xinxin Liu, Lin Niu, and Qingshan Li. 2017. "Proteomics Analysis Reveals an Important Role for the PPAR Signaling Pathway in DBDCT-Induced Hepatotoxicity Mechanisms" Molecules 22, no. 7: 1113. https://doi.org/10.3390/molecules22071113