Proteomic Analysis of Salt-Induced Changes in Protein Expression in PPARα Null Mice


PPARs are ligand-activated nuclear transcription factors that regulate β-oxidation of fatty acids in the cardiovascular system and PPARα isoform is a putative target for regulation of cardiovascular function. High salt diet is an injurious stimulus to cardiovascular function but its effect on PPARα and PPARα–associated profile of proteins is unknown. Quantitative proteomics involving a two-dimensional electrophoresis (2D-DIGE) followed by LC-MS/MS technology was used to characterize the changes in protein expression profile in the kidney, heart, and blood vessels from PPARα null (KO) and wild type (WT) mice placed on normal (0.3%, NS) or high salt (4% NaCl, HS) diet. Initial biological variation analysis using DeCyder software (v. 6.0) revealed the presence of 20 upregulated proteins and 9 proteins that are downregulated in the kidney, aorta, and heart tissues from KO and WT mice. A multimodality comparison of the differentially expressed proteins showing ≥ 1.5-fold change, ≥20% appearance at P ≤ 0.05 between strains (WT vs KO) and treatment (NS vs HS) revealed that HS diet affected 20 proteins in WT mice and 17 proteins in KO mice. However, 9 proteins were altered between WT and KO placed on NS and 7 proteins were altered by HS between WT and KO mice. The identified proteins include but not limited to those involved in fatty acid oxidation (FAO), mitochondrial electron transport chain, amino acid metabolism, stress response, DNA synthesis, and programmed cell death. HS diet led to upregulation of FAO enzymes viz: acyl-coenzyme A dehydrogenase, transketolase, and electron-transferring-flavoprotein dehydrogenase to different extents in WT and KO mice. These data showed differential and protein-specific responses to HS diet in PPARα WT and KO mice that probably reflect the functional capacities of PPARα as a means to limiting any salt-induced injury to the heart, kidney, and blood vessels.

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Obih, P. and Oyekan, A. (2014) Proteomic Analysis of Salt-Induced Changes in Protein Expression in PPARα Null Mice. Pharmacology & Pharmacy, 5, 996-1005. doi: 10.4236/pp.2014.511111.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Lemberger, T., Desvergne, B. and Wahli, W. (1996) Peroxisome Proliferator Activated Receptors: A Nuclear Receptor Signaling Pathway in Lipid Physiology. Annual Review of Cell and Developmental Biology, 12, 335-363.
[2] Bishop-Bailey, D. (2000) Peroxisome Proliferator-Activated Receptors in the Cardiovascular System. British Journal of Pharmacology, 129, 823-834.
[3] Yang, T., Michele, D.E., Park, J., Smart, A.M., Lin, Z., Brosius III, F.C., Schnermann, J.B. and Briggs, J.P. (1999) Expression of Peroxisomal Proliferator-Activated Receptors and Retinoid X Receptors in the Kidney. American Journal of Physiology, 277, 966-973.
[4] Guan, J. (2002) Targeting Peroxisome Proliferator-Activated Receptors (PPARα) in Kidney and Urologic Disease. Minerva Urologica e Nefrologica, 54, 65-79.
[5] Guan, Y., Zhang, Y., Davis, L. and Breyer, M.D. (1997) Expression of Peroxisome Proliferator-Activated Receptors in Urinary Tract of Rabbits and Humans. American Journal of Physiology, 273, 1013-1022.
[6] Djouadi, F. and Bastin, J. (2001) PPARα Gene Expression in the Developing Rat Kidney: Role of Glucocorticoids. Journal of the American Society of Nephrology, 12, 1197-1203.
[7] Hashimoto, T. (1999) Peroxisomal β-Oxidation Enzymes. Neurochemical Research, 24, 551-563.
[8] Reddy, J.K. and Mannaerts, G.P. (1994) Peroxisomal Lipid Metabolism. Annual Review of Nutrition, 14, 343-370.
[9] Aoyama, T., Peters, J.M., Iritani, N., Nakajima, T., Furihata, K., Hashimoto, T. and Gonzalez, F.J. (1998) Altered Constitutive Expression of Fatty Acid-Metabolizing Enzymes in Mice Lacking the Peroxisome Proliferator-Activated Receptor Alpha (PPAR). Journal of Biological Chemistry, 273, 5678-5684.
[10] Hashimoto, T., Fujita, T., Usuda, N., Cook, W., Qi, C., Peters, J.M., Gonzalez, F.J., Yeldandi, A.V., Rao, M.S. and Reddy, J.K. (1999) Peroxisomal and Mitochondrial Fatty Acid Oxidation in Mice Nullizygous for Both Peroxisome Proliferator-Activated Receptor and Peroxisomal Fatty Acyl-CoA Oxidase. Journal of Biological Chemistry, 274, 19228-19236.
[11] Ockner, R.K., Kaikaus, R. and Bass, N.M. (1993) Fatty-Acid Metabolism and the Pathogenesis of Hepatocellular Carcinoma: Review and Hypothesis. Hepatology, 18, 669-676.
[12] Formenty, B. and Pessayre, D. (1995) Inhibition of Mitochondrial Beta-Oxidation as a Mechanism of Hepatotoxicity. Pharmacology & Therapeutics, 67, 101-154.
[13] Ghadiminejad, I. and Saggerson, D. (1992) Physiological State and the Sensitivity of Liver Mitochondrial Outer Membrane Carnitine Palmitoyltransferase to Malonyl-CoA. Correlations with Assay Temperature, Salt Concentration and Membrane Lipid Composition. International Journal of Biochemistry, 24, 1117-1124.
[14] Rachamim, N., Latter, H., Malinin, N., Asher, C., Wald, H. and Garty, H. (1995) Dexamethasone Enhances Expression of Mitochondrial Oxidative Phosphorylation Genes in Rat Distal Colon. American Journal of Physiology, 269, C1305-C1310.
[15] Wirthensohn, G. and Guder, W.G. (1983) Renal Lipid Metabolism. Mineral Electrolyte Metabolism, 9, 203-211.
[16] Somiari, R.I., Sullivan, A., Russell, S., Somiari, S., Hu, H., Jordan, R., George, A., Katenhusen, R., Buchowiecka, A., Arciero, C., Brzeski, H., Hooke, J. and Shriver, C. (2003) High-Throughput Proteomic Analysis of Human Infiltrating Ductal Carcinoma of the Breast. Proteomics, 3, 1863-1873.
[17] Yates, J.R.D., Eng, J.K., McCormack, A.L. and Schieltz, D. (1995) Method to Correlate Tandem Mass Spectra of Modified Peptides to Amino Acid Sequences in the Protein Database. Analytical Chemistry, 67, 1426-1436.
[18] Braissant, O., Foufelle, F., Scotto, C., Dauca, M. and Wahli, W. (1996) Differential Expression of Peroxisome Proliferator-Activated Receptors (PPARα): Tissue Distribution of PPAR-α, -β, and -γ in the Adult Rat. Endocrinology, 137, 354-366.
[19] Portilla, D., Dai, G., Peters, J.M., Gonzalez, F.J., Crew, M.D. and Proia, A.D. (2000) Etomoxir-Induced PPARα-Modulated Enzymes Protects during Acute Renal Failure. American Journal of Physiology—Renal Physiology, 278, F667-F675.
[20] Li, S., Wu, P., Yarlagadda, P., Vadjunec, N.M., Proia, A.D, Harris, R.A. and Portilla, D. (2004) PPARα Ligand Protects during Cisplatin Induced Acute Renal Failure by Preventing Inhibition of Renal FAO and PDC Activity. American Journal of Physiology-Renal Physiology, 286, F572-F580.
[21] Obih, P. and Oyekan, A.O. (2008) Regulation of Blood Pressure, Natriuresis and Renal Thiazide/Amiloride Sensitivity in PPARα Null Mice. Blood Pressure, 17, 55-63.
[22] Negishi, K., Noiri, E., Sugaya, T., Li, S., Megyesi, J., Nagothu, K. and Portilla, D. (2007) A Role of Liver Fatty Acid-Binding Protein in Cisplatin-Induced Acute Renal Failure. Kidney International, 72, 348-358.
[23] Ramsay, R.R., Steenkamp, D.J. and Husain, M. (1987) Reactions of Electron-Transfer Flavoprotein and Electron-Transfer Flavoprotein: Ubiquinone Oxidoreductase. Biochemical Journal, 241, 883-892.
[24] Hirst, J. (2005) Energy Transduction by Respiratory Complex I—An Evaluation of Current Knowledge. Biochemical Society Transactions, 33, 525-529.
[25] Tian, Z., Greene, A.S., Usa, K., Matus, I.R., Bauwens, J., Pietrusz, J.L., Cowley Jr., A.W. and Liang, M. (2008) Renal Regional Proteomes in Young Dahl Salt-Sensitive Rats. Hypertension, 51, 899-904.
[26] Grussenmeyer, T., Meili-Butz, S., Roth, V., Dieterle, T., Brink, M., Matt, P., Carrel, T.P., Eckstein, F.S., Lefkpovits, I. and Grapow, M.T. (2011) Proteome Analysis in Cardiovascular Pathology Using Dahl Rat Model. Journal of Proteomics, 74, 672-682.
[27] Zheleznova, N.N., Yang, C., Ryan, R.P., Halligan, B.D., Liang, M., Greene, A.S. and Cowley Jr., A.W. (2012) Mitochondrial Proteomic Analysis Reveals Deficiencies in Oxygen Utilization in Medullary Thick Ascending Limb of Henle in Dahl Salt-Sensitive Rat. Physiological Genomics, 44, 839-842.
[28] Cruz-Topete, D., List, E.O., Okada, S., Kaider, B. and Kopchick, J.J. (2012) Proteomic Changes in the Heart of Diet-Induced Pre-Diabetic Mice. Journal of Proteomics, 74, 716-727.

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