BACKGROUND: Cachexia worsens long-term prognosis of patients with heart failure (HF). Effective treatment of cachexia is missing. We seek to characterize mechanisms of cachexia in adipose tissue, which could serve as novel targets for the treatment. METHODS: The study was conducted in advanced HF patients (n = 52; 83% male patients) undergoing heart transplantation. Patients with ≥7.5% non-intentional body weight (BW) loss during the last 6 months were rated cachectic. Clinical characteristics and circulating markers were compared between cachectic (n = 17) and the remaining, BW-stable patients. In epicardial adipose tissue (EAT), expression of selected genes was evaluated, and a combined metabolomic/lipidomic analysis was performed to assess (i) the role of adipose tissue metabolism in the development of cachexia and (ii) potential impact of cachexia-associated changes on EAT-myocardium environment. RESULTS: Cachectic vs. BW-stable patients had higher plasma levels of natriuretic peptide B (BNP; 2007 ± 1229 vs. 1411 ± 1272 pg/mL; P = 0.010) and lower EAT thickness (2.1 ± 0.8 vs. 2.9 ± 1.4 mm; P = 0.010), and they were treated with ~2.5-fold lower dose of both β-blockers and angiotensin-converting enzyme inhibitors or angiotensin receptor blockers (ACE/ARB-inhibitors). The overall pattern of EAT gene expression suggested simultaneous activation of lipolysis and lipogenesis in cachexia. Lower ratio between expression levels of natriuretic peptide receptors C and A was observed in cachectic vs. BW-stable patients (0.47 vs. 1.30), supporting activation of EAT lipolysis by natriuretic peptides. Fundamental differences in metabolome/lipidome between BW-stable and cachectic patients were found. Mitochondrial phospholipid cardiolipin (CL), specifically the least abundant CL 70:6 species (containing C16:1, C18:1, and C18:2 acyls), was the most discriminating analyte (partial least squares discriminant analysis; variable importance in projection score = 4). Its EAT levels were higher in cachectic as compared with BW-stable patients and correlated with the degree of BW loss during the last 6 months (r = -0.94; P = 0.036). CONCLUSIONS: Our results suggest that (i) BNP signalling contributes to changes in EAT metabolism in cardiac cachexia and (ii) maintenance of stable BW and 'healthy' EAT-myocardium microenvironment depends on the ability to tolerate higher doses of both ACE/ARB inhibitors and β-adrenergic blockers. In line with preclinical studies, we show for the first time in humans the association of cachexia with increased adipose tissue levels of CL. Specifically, CL 70:6 could precipitate wasting of adipose tissue, and thus, it could represent a therapeutic target to ameliorate cachexia.

• Keywords
• Adipose tissue, Cardiac cachexia, Cardiolipin, Heart failure, Lipolysis, Natriuretic peptides,
• MeSH
• Angiotensin Receptor Antagonists MeSH
• Angiotensin-Converting Enzyme Inhibitors MeSH
• Cachexia * etiology   MeSH
• Cardiolipins MeSH
• Humans MeSH
• Natriuretic Peptides MeSH
• Heart Failure * complications   MeSH
• Adipose Tissue * MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Angiotensin Receptor Antagonists MeSH
• Angiotensin-Converting Enzyme Inhibitors MeSH
• Cardiolipins MeSH
• Natriuretic Peptides MeSH

In aging, the capacity of subcutaneous adipose tissue (SAT) to store lipids decreases and this results in metabolically unfavorable fat redistribution. Triggers of this age-related SAT dysfunction may include cellular senescence or endoplasmic reticulum (ER) stress. Therefore, we compared lipogenic capacity of SAT between young and older women and investigated its relation to senescence and ER stress markers. Samples of SAT and corresponding SAT-derived primary preadipocytes were obtained from two groups of women differing in age (36 vs. 72 years, n = 15 each) but matched for fat mass. mRNA levels of selected genes (lipogenesis: ACACA, FASN, SCD1, DGAT2, ELOVL6; senescence: p16, p21, NOX4, GDF15; ER stress-ATF4, XBP1s, PERK, HSPA5, GADD34, HYOU1, CHOP, EDEM1, DNAJC3) were assessed by qPCR, protein levels of GDF15 by ELISA, and mitochondrial function by the Seahorse Analyzer. Compared to the young, SAT and in vitro differentiated adipocytes from older women exhibited reduced mRNA expression of lipogenic enzymes. Out of analyzed senescence and ER stress markers, the only gene, whose expression correlated negatively with the expression of lipogenic enzymes in both SAT and adipocytes, was GDF15, a marker of not only senescence but also mitochondrial dysfunction. In line with this, inhibition of mitochondrial ATP synthase in adipocytes strongly upregulated GDF15 while reduced expression of lipogenic enzymes. Moreover, adipocytes from older women had a tendency for diminished mitochondrial capacity. Thus, a reduced lipogenic capacity of adipocytes in aged SAT appears to be linked to mitochondrial dysfunction rather than to ER stress or accumulation of senescent cells.

• Keywords
• Aging, Lipogenesis, Mitochondrial dysfunction, Senescence, Stress of endoplasmic reticulum, Subcutaneous adipose tissue,
• MeSH
• Biomarkers metabolism   MeSH
• Cell Differentiation MeSH
• Endoplasmic Reticulum Chaperone BiP MeSH
• Adult MeSH
• Humans MeSH
• Lipogenesis * MeSH
• Mitochondria metabolism   MeSH
• Subcutaneous Fat metabolism   MeSH
• Growth Differentiation Factor 15 metabolism   MeSH
• Aged MeSH
• Cellular Senescence MeSH
• Aging metabolism   MeSH
• Endoplasmic Reticulum Stress MeSH
• Adipocytes metabolism   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Names of Substances
• Biomarkers MeSH
• Endoplasmic Reticulum Chaperone BiP MeSH
• GDF15 protein, human MeSH Browser
• HSPA5 protein, human MeSH Browser
• Growth Differentiation Factor 15 MeSH

AIMS/HYPOTHESIS: Calorie restriction is an essential component in the treatment of obesity and associated diseases. Long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) act as natural hypolipidaemics, reduce the risk of cardiovascular disease and could prevent the development of obesity and insulin resistance. We aimed to characterise the effectiveness and underlying mechanisms of the combination treatment with LC n-3 PUFA and 10% calorie restriction in the prevention of obesity and associated disorders in mice. METHODS: Male mice (C57BL/6J) were habituated to a corn-oil-based high-fat diet (cHF) for 2 weeks and then randomly assigned to various dietary treatments for 5 weeks or 15 weeks: (1) cHF, ad libitum; (2) cHF with LC n-3 PUFA concentrate replacing 15% (wt/wt) of dietary lipids (cHF + F), ad libitum; (3) cHF with calorie restriction (CR; cHF + CR); and (4) cHF + F + CR. Mice fed a chow diet were also studied. RESULTS: We show that white adipose tissue plays an active role in the amelioration of obesity and the improvement of glucose homeostasis by combining LC n-3 PUFA intake and calorie restriction in cHF-fed mice. Specifically in the epididymal fat in the abdomen, but not in other fat depots, synergistic induction of mitochondrial oxidative capacity and lipid catabolism was observed, resulting in increased oxidation of metabolic fuels in the absence of mitochondrial uncoupling, while low-grade inflammation was suppressed, reflecting changes in tissue levels of anti-inflammatory lipid mediators, namely 15-deoxy-Δ(12,15)-prostaglandin J(2) and protectin D1. CONCLUSIONS/INTERPRETATION: White adipose tissue metabolism linked to its inflammatory status in obesity could be modulated by combination treatment using calorie restriction and dietary LC n-3 PUFA to improve therapeutic strategies for metabolic syndrome.

• MeSH
• Adipose Tissue, White drug effects   metabolism   MeSH
• Diet, High-Fat MeSH
• Dietary Fats pharmacology   MeSH
• Energy Metabolism drug effects   MeSH
• Immunohistochemistry MeSH
• Caloric Restriction * MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Docosahexaenoic Acids metabolism   MeSH
• Lipid Metabolism drug effects   MeSH
• Mice, Obese MeSH
• Mice MeSH
• Fatty Acids, Omega-3 pharmacology   MeSH
• Prostaglandin D2 analogs & derivatives   metabolism   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• 9-deoxy-delta-9-prostaglandin D2 MeSH Browser
• Dietary Fats MeSH
• Docosahexaenoic Acids MeSH
• Fatty Acids, Omega-3 MeSH
• Prostaglandin D2 MeSH
• protectin D1 MeSH Browser

AIMS/HYPOTHESIS: Intake of n-3 polyunsaturated fatty acids reduces adipose tissue mass, preferentially in the abdomen. The more pronounced effect of marine-derived eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids on adiposity, compared with their precursor alpha-linolenic acid, may be mediated by changes in gene expression and metabolism in white fat. METHODS: The effects of EPA/DHA concentrate (6% EPA, 51% DHA) admixed to form two types of high-fat diet were studied in C57BL/6J mice. Oligonucleotide microarrays, cDNA PCR subtraction and quantitative real-time RT-PCR were used to characterise gene expression. Mitochondrial proteins were quantified using immunoblots. Fatty acid oxidation and synthesis were measured in adipose tissue fragments. RESULTS: Expression screens revealed upregulation of genes for mitochondrial proteins, predominantly in epididymal fat when EPA/DHA concentrate was admixed to a semisynthetic high-fat diet rich in alpha-linolenic acid. This was associated with a three-fold stimulation of the expression of genes encoding regulatory factors for mitochondrial biogenesis and oxidative metabolism (peroxisome proliferator-activated receptor gamma coactivator 1 alpha [Ppargc1a, also known as Pgc1alpha] and nuclear respiratory factor-1 [Nrf1] respectively). Expression of genes for carnitine palmitoyltransferase 1A and fatty acid oxidation was increased in epididymal but not subcutaneous fat. In the former depot, lipogenesis was depressed. Similar changes in adipose gene expression were detected after replacement of as little as 15% of lipids in the composite high-fat diet with EPA/DHA concentrate, while the development of obesity was reduced. The expression of Ppargc1a and Nrf1 was also stimulated by n-3 polyunsaturated fatty acids in 3T3-L1 cells. CONCLUSIONS/INTERPRETATION: The anti-adipogenic effect of EPA/DHA may involve a metabolic switch in adipocytes that includes enhancement of beta-oxidation and upregulation of mitochondrial biogenesis.

• MeSH
• Epididymis drug effects   metabolism   MeSH
• NF-E2-Related Factor 1 drug effects   genetics   MeSH
• Carnitine O-Palmitoyltransferase drug effects   genetics   MeSH
• Cells, Cultured MeSH
• alpha-Linolenic Acid pharmacology   MeSH
• Eicosapentaenoic Acid pharmacology   MeSH
• Docosahexaenoic Acids pharmacology   MeSH
• Lipogenesis drug effects   MeSH
• Mitochondrial Proteins drug effects   metabolism   MeSH
• Mitochondria drug effects   metabolism   MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Fatty Acids, Unsaturated isolation & purification   metabolism   pharmacology   MeSH
• Obesity prevention & control   MeSH
• Oxidation-Reduction MeSH
• Subcutaneous Fat drug effects   metabolism   MeSH
• Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha MeSH
• Gene Expression Regulation drug effects   MeSH
• Fish Oils chemistry   MeSH
• Trans-Activators drug effects   genetics   MeSH
• Transcription Factors MeSH
• Adipose Tissue drug effects   metabolism   MeSH
• Adipocytes drug effects   metabolism   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Names of Substances
• NF-E2-Related Factor 1 MeSH
• Carnitine O-Palmitoyltransferase MeSH
• alpha-Linolenic Acid MeSH
• Eicosapentaenoic Acid MeSH
• Docosahexaenoic Acids MeSH
• Mitochondrial Proteins MeSH
• Fatty Acids, Unsaturated MeSH
• Ppargc1a protein, mouse MeSH Browser
• Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha MeSH
• Fish Oils MeSH
• Trans-Activators MeSH
• Transcription Factors MeSH

In vitro experiments suggest that stimulation of lipolysis by catecholamines in adipocytes depends on the energy status of these cells. We tested whether mitochondrial uncoupling proteins (UCPs) that control the efficiency of ATP production could affect lipolysis and noradrenaline signalling in white fat in vivo. The lipolytic effect of noradrenaline was lowered by ectopic UCP1 in white adipocytes of aP2-Ucp1 transgenic mice, overexpressing the UCP1 gene from the aP2 gene promoter, reflecting the magnitude of UCP1 expression, the impaired stimulation of cAMP levels by noradrenaline and the reduction of the ATP/ADP ratio in different fat depots. Thus only subcutaneous but not epididymal fat was affected. UCP1 also down-regulated the expression of hormone-sensitive lipase and lowered its activity, and altered the expression of trimeric G-proteins in adipocytes. The adipose tissue content of the stimulatory G-protein alpha subunit was increased while that of the inhibitory G-protein alpha subunits decreased in response to UCP1 expression. Our results support the idea that the energy status of cells, and the ATP/ADP ratio in particular, modulates the lipolytic effects of noradrenaline in adipose tissue in vivo. They also demonstrate changes at the G-protein level that tend to overcome the reduction of lipolysis when ATP level in adipocytes is low. Therefore, respiratory uncoupling may exert a broad effect on hormonal signalling in adipocytes.

• MeSH
• Adenosine Diphosphate metabolism   MeSH
• Adenosine Triphosphate metabolism   MeSH
• Cyclic AMP metabolism   MeSH
• DNA Primers MeSH
• Genotype MeSH
• Hepatocytes enzymology   MeSH
• Ion Channels MeSH
• Lipase genetics   MeSH
• Lipolysis * MeSH
• Membrane Proteins genetics   physiology   MeSH
• Mitochondrial Proteins MeSH
• Mice, Inbred C57BL MeSH
• Mice, Transgenic MeSH
• Mice MeSH
• Norepinephrine metabolism   MeSH
• GTP-Binding Proteins metabolism   MeSH
• Gene Expression Regulation, Enzymologic MeSH
• Base Sequence MeSH
• Carrier Proteins genetics   physiology   MeSH
• Adipose Tissue enzymology   metabolism   MeSH
• Uncoupling Protein 1 MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Adenosine Diphosphate MeSH
• Adenosine Triphosphate MeSH
• Cyclic AMP MeSH
• DNA Primers MeSH
• Ion Channels MeSH
• Lipase MeSH
• Membrane Proteins MeSH
• Mitochondrial Proteins MeSH
• Norepinephrine MeSH
• GTP-Binding Proteins MeSH
• Carrier Proteins MeSH
• Ucp1 protein, mouse MeSH Browser
• Uncoupling Protein 1 MeSH