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TIMP3 interplays with apelin to regulate cardiovascular metabolism in hypercholesterolemic mice.

Stöhr R, Kappel BA, Carnevale D, Cavalera M, Mavilio M, Arisi I, Fardella V, Cifelli G, Casagrande V, Rizza S, Cattaneo A, Mauriello A, Menghini R, Lembo G, Federici M - Mol Metab (2015)

Bottom Line: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan.Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids.Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, Italy ; Department of Internal Medicine I, University Hospital Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.

ABSTRACT

Objective: Tissue inhibitor of metalloproteinase 3 (TIMP3) is an extracellular matrix (ECM) bound protein, which has been shown to be downregulated in human subjects and experimental models with cardiometabolic disorders, including type 2 diabetes mellitus, hypertension and atherosclerosis. The aim of this study was to investigate the effects of TIMP3 on cardiac energy homeostasis during increased metabolic stress conditions.

Methods: ApoE(-/-)TIMP3(-/-) and ApoE(-/-) mice on a C57BL/6 background were subjected to telemetric ECG analysis and experimental myocardial infarction as models of cardiac stress induction. We used Western blot, qRT-PCR, histology, metabolomics, RNA-sequencing and in vivo phenotypical analysis to investigate the molecular mechanisms of altered cardiac energy metabolism.

Results: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan. Telemetric ECG analysis showed increased arrhythmic episodes, and experimental myocardial infarction by left anterior descending artery (LAD) ligation resulted in increased peri-operative mortality together with increased scar formation, ventricular dilatation and a reduction of cardiac function after 4 weeks in the few survivors. Hearts of ApoE(-/-)TIMP3(-/-) exhibited accumulation of neutral lipids when fed a chow diet, which was exacerbated by a high fat, high cholesterol diet. Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids. Using whole heart mRNA sequencing, we identified apelin as a putative modulator of these metabolic defects. Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice. Injection of apelin restored the hitherto identified metabolic defects of lipid oxidation.

Conclusion: TIMP3 regulates lipid metabolism as well as oxidative stress response via apelin. These findings therefore suggest that TIMP3 maintains metabolic flexibility in the heart, particularly during episodes of increased cardiac stress.

No MeSH data available.


Related in: MedlinePlus

ApoE−/−TIMP3−/− mice have impairment of fatty acid metabolism. A) Metabolomics analysis of the serum shows a decrease in long-chain monounsaturated FFAs, medium-chain acyl-carnitines and medium-chain fatty acid markers in ApoE−/−TIMP3−/− animals (*p ≤ 0.05 and **p < 0.01, Student's t test, n = 6 per group). B) Regulation of MUFA/SUFA balance in mammalian cell lipids by SCD1.
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fig4: ApoE−/−TIMP3−/− mice have impairment of fatty acid metabolism. A) Metabolomics analysis of the serum shows a decrease in long-chain monounsaturated FFAs, medium-chain acyl-carnitines and medium-chain fatty acid markers in ApoE−/−TIMP3−/− animals (*p ≤ 0.05 and **p < 0.01, Student's t test, n = 6 per group). B) Regulation of MUFA/SUFA balance in mammalian cell lipids by SCD1.

Mentions: To unveil biomarkers associated to the reduced lifespan and impaired lipid oxidation, we performed metabolomics analysis in ApoE−/− and ApoE−/−TIMP3−/−. Random Forest analysis of the analyzed biochemicals resulted in a predictive accuracy of 83% in classifying between ApoE−/− and ApoE−/−TIMP3−/− mice (Supplementary Figure S3). Several metabolites were differently regulated in the ApoE−/−TIMP3−/− compared with ApoE−/− littermates (Supplementary Tables S1 and S2). In the serum of ApoE−/−TIMP3−/− mice, we observed complex modulation of pathways linked to oxidative stress as demonstrated by the depletion of ergothioneine, equol sulfate, cysteine-glutathione disulfide and oxidized glutathione while 2-hydroxybutyrate and 2-aminobutyrate were increased (Figure 4A,B). ApoE−/−TIMP3−/− mice also showed reductions in long-chain monounsaturated FFAs such as 10-nonadecenoate and 10-heptadecenoate, medium-chain acyl-carnitines such as hexanoylcarnitine, medium-chain acyl-glycine (i.e., valerylglycine) and the medium-chain fatty acid marker of beta-oxidation, 3-hydroxydecanoate (Figure 5A,B). On the other hand, the long-chain acyl-carnitine stearoylcarnitine was significantly elevated in ApoE−/−TIMP3−/− mice (Figure 5A,B). ApoE−/−TIMP3−/− mice also showed altered sphingolipid metabolism with increased levels of palmitoyl sphingomyelin, stearoyl sphingomyelin, 1-stearoylglycerol and sphingosin that can be in part connected to increased stearoylcarnitine and its derivatives (Figure 6A,B).


TIMP3 interplays with apelin to regulate cardiovascular metabolism in hypercholesterolemic mice.

Stöhr R, Kappel BA, Carnevale D, Cavalera M, Mavilio M, Arisi I, Fardella V, Cifelli G, Casagrande V, Rizza S, Cattaneo A, Mauriello A, Menghini R, Lembo G, Federici M - Mol Metab (2015)

ApoE−/−TIMP3−/− mice have impairment of fatty acid metabolism. A) Metabolomics analysis of the serum shows a decrease in long-chain monounsaturated FFAs, medium-chain acyl-carnitines and medium-chain fatty acid markers in ApoE−/−TIMP3−/− animals (*p ≤ 0.05 and **p < 0.01, Student's t test, n = 6 per group). B) Regulation of MUFA/SUFA balance in mammalian cell lipids by SCD1.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4588459&req=5

fig4: ApoE−/−TIMP3−/− mice have impairment of fatty acid metabolism. A) Metabolomics analysis of the serum shows a decrease in long-chain monounsaturated FFAs, medium-chain acyl-carnitines and medium-chain fatty acid markers in ApoE−/−TIMP3−/− animals (*p ≤ 0.05 and **p < 0.01, Student's t test, n = 6 per group). B) Regulation of MUFA/SUFA balance in mammalian cell lipids by SCD1.
Mentions: To unveil biomarkers associated to the reduced lifespan and impaired lipid oxidation, we performed metabolomics analysis in ApoE−/− and ApoE−/−TIMP3−/−. Random Forest analysis of the analyzed biochemicals resulted in a predictive accuracy of 83% in classifying between ApoE−/− and ApoE−/−TIMP3−/− mice (Supplementary Figure S3). Several metabolites were differently regulated in the ApoE−/−TIMP3−/− compared with ApoE−/− littermates (Supplementary Tables S1 and S2). In the serum of ApoE−/−TIMP3−/− mice, we observed complex modulation of pathways linked to oxidative stress as demonstrated by the depletion of ergothioneine, equol sulfate, cysteine-glutathione disulfide and oxidized glutathione while 2-hydroxybutyrate and 2-aminobutyrate were increased (Figure 4A,B). ApoE−/−TIMP3−/− mice also showed reductions in long-chain monounsaturated FFAs such as 10-nonadecenoate and 10-heptadecenoate, medium-chain acyl-carnitines such as hexanoylcarnitine, medium-chain acyl-glycine (i.e., valerylglycine) and the medium-chain fatty acid marker of beta-oxidation, 3-hydroxydecanoate (Figure 5A,B). On the other hand, the long-chain acyl-carnitine stearoylcarnitine was significantly elevated in ApoE−/−TIMP3−/− mice (Figure 5A,B). ApoE−/−TIMP3−/− mice also showed altered sphingolipid metabolism with increased levels of palmitoyl sphingomyelin, stearoyl sphingomyelin, 1-stearoylglycerol and sphingosin that can be in part connected to increased stearoylcarnitine and its derivatives (Figure 6A,B).

Bottom Line: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan.Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids.Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, Italy ; Department of Internal Medicine I, University Hospital Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.

ABSTRACT

Objective: Tissue inhibitor of metalloproteinase 3 (TIMP3) is an extracellular matrix (ECM) bound protein, which has been shown to be downregulated in human subjects and experimental models with cardiometabolic disorders, including type 2 diabetes mellitus, hypertension and atherosclerosis. The aim of this study was to investigate the effects of TIMP3 on cardiac energy homeostasis during increased metabolic stress conditions.

Methods: ApoE(-/-)TIMP3(-/-) and ApoE(-/-) mice on a C57BL/6 background were subjected to telemetric ECG analysis and experimental myocardial infarction as models of cardiac stress induction. We used Western blot, qRT-PCR, histology, metabolomics, RNA-sequencing and in vivo phenotypical analysis to investigate the molecular mechanisms of altered cardiac energy metabolism.

Results: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan. Telemetric ECG analysis showed increased arrhythmic episodes, and experimental myocardial infarction by left anterior descending artery (LAD) ligation resulted in increased peri-operative mortality together with increased scar formation, ventricular dilatation and a reduction of cardiac function after 4 weeks in the few survivors. Hearts of ApoE(-/-)TIMP3(-/-) exhibited accumulation of neutral lipids when fed a chow diet, which was exacerbated by a high fat, high cholesterol diet. Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids. Using whole heart mRNA sequencing, we identified apelin as a putative modulator of these metabolic defects. Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice. Injection of apelin restored the hitherto identified metabolic defects of lipid oxidation.

Conclusion: TIMP3 regulates lipid metabolism as well as oxidative stress response via apelin. These findings therefore suggest that TIMP3 maintains metabolic flexibility in the heart, particularly during episodes of increased cardiac stress.

No MeSH data available.


Related in: MedlinePlus