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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−/− animals reveal decreased lifespan, arrhythmias and susceptibility to increased cardiac stress. A) Combined retro- and prospective survival analysis in a cohort of 64 male and female mice (n = 32 for ApoE−/−TIMP3−/− M/F = 12/20 and n = 32 for ApoE−/− M/F = 12/20) over a time period of 12 months (p = 0.0002, Cox Test). B) Telemetry 24 h ECG in 32-weeks-old ApoE−/−TIMP3−/− and ApoE−/− mice. We observed a higher amount of arrhythmic episodes in ApoE−/−TIMP3−/− as well as sudden deaths in 4 out of 6 male mice during the 7-day registration (p = 0.06, Fisher's exact test). C) Increased peri-operative mortality during experimental myocardial infarction by LAD ligation in ApoE−/−TIMP3−/− (13/19) compared to ApoE−/− (3/13) (***p < 0.001 by Chi-square test). D) Representative photomicrographs of Masson-Trichrome stained hearts 4 weeks post-LAD ligation showing increased infarction size as confirmed by morphometric quantification of percent total LV circumference. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 3–4 per group). E) Reduced systolic (dp/dtmax) and diastolic (dp/dtmin) heart function in ApoE−/−TIMP3−/− mice 4 weeks after LAD ligation as measured by Millar Catheter. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 4–5 per group).
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fig1: ApoE−/−TIMP3−/− animals reveal decreased lifespan, arrhythmias and susceptibility to increased cardiac stress. A) Combined retro- and prospective survival analysis in a cohort of 64 male and female mice (n = 32 for ApoE−/−TIMP3−/− M/F = 12/20 and n = 32 for ApoE−/− M/F = 12/20) over a time period of 12 months (p = 0.0002, Cox Test). B) Telemetry 24 h ECG in 32-weeks-old ApoE−/−TIMP3−/− and ApoE−/− mice. We observed a higher amount of arrhythmic episodes in ApoE−/−TIMP3−/− as well as sudden deaths in 4 out of 6 male mice during the 7-day registration (p = 0.06, Fisher's exact test). C) Increased peri-operative mortality during experimental myocardial infarction by LAD ligation in ApoE−/−TIMP3−/− (13/19) compared to ApoE−/− (3/13) (***p < 0.001 by Chi-square test). D) Representative photomicrographs of Masson-Trichrome stained hearts 4 weeks post-LAD ligation showing increased infarction size as confirmed by morphometric quantification of percent total LV circumference. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 3–4 per group). E) Reduced systolic (dp/dtmax) and diastolic (dp/dtmin) heart function in ApoE−/−TIMP3−/− mice 4 weeks after LAD ligation as measured by Millar Catheter. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 4–5 per group).

Mentions: A combined retro- and prospective 52 weeks analysis of ApoE−/−TIMP3−/− (n = 32) mice revealed a decreased survival rate when compared to ApoE−/− (n = 32) littermates (p = 0.0002, Cox Test; Figure 1A), with the first episodes of death observed at week 28 in the ApoE−/−TIMP3−/− group. The higher mortality rate in the ApoE−/−TIMP3−/− group was independent from gender and no differences between genotypes for specific macroscopic signs of infections including dermatitis or sudden loss of body weight were observed. Since necroscopy of ApoE−/−TIMP3−/− mice did not reveal any obvious sign such as bleeding, malignancy or macroscopically visible organ pathology responsible for decreased lifespan, we speculated that mice might suffer from sudden cardiac death.


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−/− animals reveal decreased lifespan, arrhythmias and susceptibility to increased cardiac stress. A) Combined retro- and prospective survival analysis in a cohort of 64 male and female mice (n = 32 for ApoE−/−TIMP3−/− M/F = 12/20 and n = 32 for ApoE−/− M/F = 12/20) over a time period of 12 months (p = 0.0002, Cox Test). B) Telemetry 24 h ECG in 32-weeks-old ApoE−/−TIMP3−/− and ApoE−/− mice. We observed a higher amount of arrhythmic episodes in ApoE−/−TIMP3−/− as well as sudden deaths in 4 out of 6 male mice during the 7-day registration (p = 0.06, Fisher's exact test). C) Increased peri-operative mortality during experimental myocardial infarction by LAD ligation in ApoE−/−TIMP3−/− (13/19) compared to ApoE−/− (3/13) (***p < 0.001 by Chi-square test). D) Representative photomicrographs of Masson-Trichrome stained hearts 4 weeks post-LAD ligation showing increased infarction size as confirmed by morphometric quantification of percent total LV circumference. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 3–4 per group). E) Reduced systolic (dp/dtmax) and diastolic (dp/dtmin) heart function in ApoE−/−TIMP3−/− mice 4 weeks after LAD ligation as measured by Millar Catheter. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 4–5 per group).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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

fig1: ApoE−/−TIMP3−/− animals reveal decreased lifespan, arrhythmias and susceptibility to increased cardiac stress. A) Combined retro- and prospective survival analysis in a cohort of 64 male and female mice (n = 32 for ApoE−/−TIMP3−/− M/F = 12/20 and n = 32 for ApoE−/− M/F = 12/20) over a time period of 12 months (p = 0.0002, Cox Test). B) Telemetry 24 h ECG in 32-weeks-old ApoE−/−TIMP3−/− and ApoE−/− mice. We observed a higher amount of arrhythmic episodes in ApoE−/−TIMP3−/− as well as sudden deaths in 4 out of 6 male mice during the 7-day registration (p = 0.06, Fisher's exact test). C) Increased peri-operative mortality during experimental myocardial infarction by LAD ligation in ApoE−/−TIMP3−/− (13/19) compared to ApoE−/− (3/13) (***p < 0.001 by Chi-square test). D) Representative photomicrographs of Masson-Trichrome stained hearts 4 weeks post-LAD ligation showing increased infarction size as confirmed by morphometric quantification of percent total LV circumference. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 3–4 per group). E) Reduced systolic (dp/dtmax) and diastolic (dp/dtmin) heart function in ApoE−/−TIMP3−/− mice 4 weeks after LAD ligation as measured by Millar Catheter. Data are mean ± SEM (*p < 0.05 by Student t-test, n = 4–5 per group).
Mentions: A combined retro- and prospective 52 weeks analysis of ApoE−/−TIMP3−/− (n = 32) mice revealed a decreased survival rate when compared to ApoE−/− (n = 32) littermates (p = 0.0002, Cox Test; Figure 1A), with the first episodes of death observed at week 28 in the ApoE−/−TIMP3−/− group. The higher mortality rate in the ApoE−/−TIMP3−/− group was independent from gender and no differences between genotypes for specific macroscopic signs of infections including dermatitis or sudden loss of body weight were observed. Since necroscopy of ApoE−/−TIMP3−/− mice did not reveal any obvious sign such as bleeding, malignancy or macroscopically visible organ pathology responsible for decreased lifespan, we speculated that mice might suffer from sudden cardiac death.

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