Liver ATP Research Articles

Reduced Liver Mitochondrial Energy Metabolism Impairs Food Intake Regulation Following Gastric Preloads and Fasting.

The capacity of the liver to serve as a peripheral sensor in the regulation of food intake has been debated for over half a century. The anatomical position and physiological roles of the liver suggest it is a prime candidate to serve as an interoceptive sensor of peripheral tissue and systemic energy state. Importantly, maintenance of liver ATP levels and within- meal food intake inhibition is impaired in human subjects with obesity and obese pre- clinical models. We demonstrate that decreased hepatic mitochondrial energy metabolism in liver-specific, heterozygous PGC1a mice results in reduced mitochondrial response to changes in ΔGATP and tissue ATP following fasting. These impairments in liver energy state are associated with larger and longer meals during chow feeding, impaired dose-dependent food intake inhibition in response to mixed and individual nutrient oral pre-loads, and greater acute fasting-induced food intake. These data support previous work proposing liver-mediated food intake regulation through modulation of peripheral satiation signals.

Characterization of glucose metabolism in high-growth performance Nile tilapia (Oreochromis niloticus)

Significant differences in growth performance have been observed in fish that have undergone the same culture cycle, even under identical conditions, including genetic background, water environment, and feeding strategy. These differences may adversely affect the harvesting and subsequent processing of commercial fish. We have previously demonstrated that the genes of glucose metabolism in high-growth performance tilapia (Oreochromis niloticus) showed significant upregulation at the transcriptome level. These findings suggested that the growth performance of tilapia may be closely related to their endogenous glucose metabolism capacity. To investigate the characteristics of glucose metabolism in high-growth performance tilapia, three full-sibling families were cultured under the same environmental and feeding conditions. The high-growth performance group (HG) and low-growth performance group (LG) were screened and compared at the biochemical and transcriptional levels during the postprandial period of 0 to 7 h. Based on biochemical analysis, HG had higher levels of intestinal lipase activity (0 to 1 h), serum triglyceride content (0 to 7 h), liver acetyl coenzyme A content (1 to 4 h), and liver glycogen content (0 to 1 h and 6 to 7 h) than LG. In contrast, HG had lower serum glucose content (0 h and 2 to 4 h), ATP content (4 to 7 h), and liver pyruvate content (3 to 4 h) than LG. There were no significant differences between HG and LG in liver ATP, intestinal α-amylase, and trypsin. We conducted a comprehensive analysis of gene expression within multiple metabolic pathways, namely the tricarboxylic acid cycle, glycolysis, pentose phosphate, glycogen metabolism, gluconeogenesis, and insulin signaling pathways, during three specific post-feeding periods: 0 to 1 h, 2–5 h, and 6 to 7 h, and revealed unique gene expression patterns related to glucose metabolic pathways in HG at different stages of post-feeding. We first revealed the endogenous glucose metabolism characteristics of high-growth performance tilapia. The findings will provide novel insights for further research on quality improvement related to growth performance in fish.

Effects of Shenling Baizhu powder on intestinal microflora metabolites and liver mitochondrial energy metabolism in nonalcoholic fatty liver mice.

Non-alcoholic fatty liver disease (NAFLD) is characterised by the excessive accumulation of triglycerides in the liver. Shenling Baizhu powder (SLBZP) is formulated from various natural medicinal plants that protect the liver and are used to treat intestinal diseases. SLBZP improves the symptoms of NAFLD. However, its mechanism of action remains unclear. Herein, we investigated the ameliorative effect of SLBZP on model mice with high-fat-diet (HFD)-induced NAFLD. Additionally, we evaluated the impact of SLBZP on the intestinal flora and its metabolites and mitochondrial energy metabolism in NAFLD. We used HFD to establish a mouse model of NAFLD. Different drug interventions were administered. We measured serum biochemical indices. Liver sections were visualised with hematoxylin-eosin and oil red O staining. 16S rDNA amplicon sequencing technology was used to analyse the diversity and abundance of the intestinal flora. Short-chain fatty acids (SCFAs) in the intestinal contents were detected using GC-MS. Liver tissue was sampled to detect mitochondrial membrane functional indices. Western blotting was used to determine the levels of mitochondrial pathway-related proteins, namely, uncoupling protein 2 (UCP2), adenosine monophosphate-activated protein kinase (AMPK) and inhibitory factor 1 (IF1) of F1Fo ATP synthesis/hydrolase, in the liver. The spleen-invigorating classic recipe of SLBZP reduced liver lipid deposition in mice with HFD-induced NAFLD. Additionally, SCFAs produced by intestinal flora metabolism regulated the UCP2/AMPK/IF1 signalling pathway involved in liver mitochondrial energy metabolism to improve the liver mitochondrial membrane permeability, respiratory state and oxidative phosphorylation efficiency of mice with NAFLD. Finally, SLBZP increased the liver ATP level. Our results suggest that the therapeutic effect of SLBZP on NAFLD is related to the regulation of hepatic mitochondrial energy metabolism by intestinal flora and its metabolites and is possibly associated with the UCP2/AMPK/IF1 signalling pathway.

Elaidic acid leads to mitochondrial dysfunction via mitochondria-associated membranes triggers disruption of mitochondrial calcium fluxes

Elaidic acid (EA) stimulation can lead to endoplasmic reticulum stress (ERS), accompanied by a large release of Ca2+, and ultimately the activation of NLRP3 inflammasome in Kupffer cells (KCs). Mitochondrial instability or dysfunction may be the key stimulating factors to activate NLRP3 inflammasome, and sustained Ca2+ transfer can result in mitochondrial dysfunction. We focused on KCs to explore the damage to mitochondria by EA. After EA stimulation, cells produced an oxidative stress (OS) response with a significant increase in ROS release. Immunoprecipitation experiments and the addition of inhibitors revealed that the increase in the level of intracellular Ca2+ led to Ca2+ accumulation in the mitochondrial matrix via mitochondria-associated membranes (MAMs). This was accompanied by a significant release of mROS, loss of MMP and ATP, and a significant increase in mitochondrial permeability transition pore opening, ultimately leading to mitochondrial instability. These findings confirmed the mechanism that EA induced mitochondrial Ca2+ imbalance in KCs via MAM, ultimately leading to mitochondrial dysfunction. Meanwhile, EA induced OS and the decrease of MMP and ATP in rat liver, and significant lesions were found in liver mitochondria. Swelling of the inner mitochondrial cristae and mitochondrial vacuolization occurred, with a marked increase in lipid droplets.

A comparative study on the effects of supplemental L-carnitine on fatty acids composition, energy status and related genes expression in Cyprinus carpio var. Jian and Rhynchocypris lagowskii (Dybowski, 1869)

This study was to compare study on the effects of supplemental L-carnitine on fatty acids composition, energy status and related genes expression in Cyprinus carpio var. Jian and Rhynchocypris lagowskii. Different concentrations of L-carnitine were added to the basal diets of two species of fish, in which Cyprinus carpio were supplemented with 0, 50 (LC), 200 (AC) and 800 (HC) mg/kg, and Rhynchocypris lagowskii were supplemented at 0, 200 (LC), 400 (AC) and 800 (HC) mg/kg, respectively, for 8 weeks. Results showed that in liver of AC group on C. carpio, the changes of HUFA and ∑DHA+EPA showed a significant downward trend, while the changes of PUFA, EFA and ∑n3 + n6 showed an upward trend. The above trend in liver of the AC group on R. lagowskii was opposite to C. carpio. With the exception of HUFA, the changes of fatty acids of AC group in muscles of the two fish were completely different. The content of ATP and AMP in liver and muscle in AC and HC groups of two fish species showed a decreasing trend, while the change trend of ADP content was opposite. AMP/ATP ratio in liver and muscle showed a trend of first increasing and then decreasing in the treatment groups of two fish species. The relative gene expression levels of AMPKα1, AMPKα2, ACC2 and CPT-I of liver and muscle in treatment groups of C. carpio showed a decreasing trend, while these values showed an opposite trend in liver of R. lagowskii. Consequently, the different fish species may be the main reason that affects the regulation of fatty acid metabolism by exogenous L-carnitine. In addition, AMPK, acting as an energy receptor is a key effector in regulation of fatty acids metabolism by L-carnitine. Further study of AMPK function in fatty acids metabolism of fish is thus warranted.

Liver energy homeostasis impacts hypothalamus gene expression and food intake regulation

Several neuronal populations within the hypothalamus are known to modulate feeding behavior in mammals. Specifically, the arcuate nucleus contains appetite‐suppressing neurons pro‐opiomelanocortin (POMC), and appetite‐stimulating neurons neuropeptide Y (NPY)/agouti‐related peptide (AgRP) neurons. These neurons receive peripheral signals through blood and peripheral vagal afferents fibers. However, it is not well understood how peripheral signals, such as liver energy metabolism, may impact these neuronal populations to modulate feeding behavior. Herein, we used a hepatocyte‐specific PGC1a heterozygous (LPGC1a) mouse model, with associated reductions in mitochondrial fatty acid oxidation and respiratory capacity, to assess the role of liver energy homeostasis in the expression of arcuate nucleus genes involved in food intake regulation, and acute feeding behavior after peripheral delivery of nutrient or peripheral satiation hormones. To assess the impact of liver energy changes on arcuate nucleus gene expression, we used a fasting/refeeding paradigm (16‐hour fast, 2‐hour food withdrawal, and 16‐hour fast with a 4‐hour refeed). We observed reductions in liver PGC1a mRNA expression in LPGC1a compared to wildtype (WT) during all fasting and refed states. LPGC1a mice had significantly lower liver ATP and increased AMP during a fasted state. Suggesting that during a fasted state, the LPGC1a mice struggle to maintain liver energy homeostasis. Furthermore, LPGC1a mice had significantly increased AgRP mRNA expression in the hypothalamus during a fast. Interestingly, POMC expression was lower in LPGC1a mice at 2hrs and refed compared to WT and showed no change across any of the energy states. Importantly, acute food intake following 2 hr food withdrawal was not inhibited by intraperitoneal glucose or GLP‐1 in LPGC1a mice. Together these results suggest that a reduced liver energy state modulates activity of hypothalamic POMC and AgRP neurons, potentially contributing to alterations in feeding behavior of LPGC1a mice previously observed.

Effects of Chronic Thermal Stress on Performance, Energy Metabolism, Antioxidant Activity, Brain Serotonin, and Blood Biochemical Indices of Broiler Chickens.

Simple SummaryIn the tropical and subtropical regions, heat stress is the main limiting factor of poultry industries. In this context, broilers are more liable to thermal stress due to their fast growth, rapid metabolic rate, and high level of production. The aim of the current work was to analyze changes in the brain serotonin, energy metabolism, antioxidant biomarkers, and blood chemistry of broiler chickens subjected to chronic thermal stress. Thermal stress disturbed the antioxidant defense system and energy metabolism and exhausted ATP levels in the liver tissues of broiler chickens. Interestingly, chronic thermal stress reduced the level of brain serotonin and the activity of CoQ10 in liver tissues.The aim of this paper was to investigate the effects of chronic thermal stress on the performance, energy metabolism, liver CoQ10, brain serotonin, and blood parameters of broiler chickens. In total, 100 one-day-old chicks were divided into two equal groups of five replicates. At 22 days of age and thereafter, the first group (TN) was maintained at a thermoneutral condition (23 ± 1 °C), while the second group (TS) was subjected to 8 h of thermal stress (34 °C). The heat-stressed group showed significantly lower ADFI but higher FCR than the thermoneutral group (p = 0.030 and 0.041, respectively). The TS group showed significantly higher serum cholesterol, ALT, and AST (p = 0.033, 0.024, and 0.010, respectively). Meanwhile, the TS group showed lower serum total proteins, albumin, globulin, and Na+ than the TN group (p = 0.001, 0.025, 0.032, and 0.002, respectively). Furthermore, the TS group showed significantly lower SOD and catalase in heart tissues (p = 0.005 and 0.001, respectively). The TS group showed significantly lower liver ATP than the TN group (p = 0.005). Meanwhile, chronic thermal stress significantly increased the levels of ADP and AMP in the liver tissues of broiler chickens (p = 0.004 and 0.029, respectively). The TS group showed significantly lower brain serotonin (p = 0.004) and liver CoQ10 (p = 0.001) than the TN group. It could be concluded that thermal stress disturbed the antioxidant defense system and energy metabolism and exhausted ATP levels in the liver tissues of broiler chickens. Interestingly, chronic thermal stress reduced the level of brain serotonin and the activity of CoQ10 in liver tissues.

Branched-chain amino acids and l-carnitine attenuate lipotoxic hepatocellular damage in rat cirrhotic liver

Branched-chain amino acids (BCAA) reverse malnutrition and l-carnitine leads to the reduction of hyperammonemia and muscle cramps in cirrhotic patients. BCAA and l-carnitine are involved in glucose and fatty acid metabolism, however their mechanistic activity in cirrhotic liver is not fully understood. We aim to define the molecular mechanism(s) and combined effects of BCAA and l-carnitine using a cirrhotic rat model. Rats were administered carbon tetrachloride for 10 weeks to induce cirrhosis. During the last 6 weeks of administration, cirrhotic rats received BCAA, l-carnitine or a combination of BCAA and l-carnitine daily via gavage. We found that BCAA and l-carnitine treatments significantly improved hepatocellular function associated with reduced triglyceride level, lipid deposition and adipophilin expression, in cirrhotic liver. Lipidomic analysis revealed dynamic changes in hepatic lipid composition by BCAA and l-carnitine administrations. BCAA and l-carnitine globally increased molecular species of phosphatidylcholine. Liver triacylglycerol and phosphatidylcholine hydroperoxides were significantly decreased by BCAA and l-carnitine. Furthermore, serum and liver ATP levels were significantly increased in all treatments, which were attributed to the elevation of mature cardiolipins and mitochondrial component gene expressions. Finally, BCAA and l-carnitine dramatically reduced hepatocellular death. In conclusion, BCAA and l-carnitine treatments attenuate hepatocellular damage through the reduction of lipid peroxides and the overall maintenance of mitochondrial integrity within the cirrhotic liver. These effectiveness of BCAA and l-carnitine support the therapeutic strategies in human chronic liver diseases.

Hepatic HuR modulates lipid homeostasis in response to high-fat diet

Lipid transport and ATP synthesis are critical for the progression of non-alcoholic fatty liver disease (NAFLD), but the underlying mechanisms are largely unknown. Here, we report that the RNA-binding protein HuR (ELAVL1) forms complexes with NAFLD-relevant transcripts. It associates with intron 24 of Apob pre-mRNA, with the 3′UTR of Uqcrb, and with the 5′UTR of Ndufb6 mRNA, thereby regulating the splicing of Apob mRNA and the translation of UQCRB and NDUFB6. Hepatocyte-specific HuR knockout reduces the expression of APOB, UQCRB, and NDUFB6 in mice, reducing liver lipid transport and ATP synthesis, and aggravating high-fat diet (HFD)-induced NAFLD. Adenovirus-mediated re-expression of HuR in hepatocytes rescues the effect of HuR knockout in HFD-induced NAFLD. Our findings highlight a critical role of HuR in regulating lipid transport and ATP synthesis.

An atlas of the catalytically active liver and spleen kinases in chicken identified by chemoproteomics

Phosphorylation is a post-translational protein modification regulating most known cellular processes. While protein kinases constitute a large family of highly conserved enzymes, identification of active kinases is challenging due to a low abundance of some of these signaling molecules. Although chicken is the first agricultural animal to have a sequenced genome, annotation of the kinome, i.e., a complement of all protein kinases in the genome is limited. We used chemical probes consisting of ATP and ADP derivatives binding to specific lysine (Lys) residues within the ATP-binding pocket of kinases, combined with proteomics, to identify 267 peptides labeled with the ATP and ADP acyl derivatives and 188 corresponding chicken kinases in chicken spleen and liver. Our description of active chicken kinases and ATP binding sites will support future studies focused on identifying the role of this important class of enzymes in chicken health and disease. SignificanceAdvances made in understanding chicken enzymes are critical for the improved knowledge of the regulatory pathways controlling physiological processes in chicken. Since protein phosphorylation controls multiple aspects of cell fate, it is often linked to pathological conditions, and understanding of the kinase expression in chicken is essential for future therapeutic approaches. We coupled proteomics and labeling with active-site probes binding to Lys residues within the ATP-binding pocket of kinases to identify 188 kinases and corresponding 267 peptides labeled with the ATP and ADP acyl derivatives in chicken spleen and liver. Results of the present study describing catalytically active kinases is a starting point for chemoproteomic-based interrogation of kinases in chicken exposed to different conditions. Kinases identified in this study are available through the Chickspress genome browser that has previously published mRNA, miRNA, and shotgun proteomics data.

Adding the oxygen carrier M101 to a cold-storage solution could be an alternative to HOPE for liver graft preservation.

Background & AimsHypothermic oxygenated machine perfusion (HOPE) is a promising technique for providing oxygen to the liver during graft preservation; however, because of associated logistical constraints, addition of an oxygen transporter to static cold-storage solutions (SCS) might be easier. M101 is marine worm haemoglobin that has been shown to improve kidney preservation in the clinic when added to SCS. This study evaluated the effects of the addition of M101 to SCS on the quality of pig liver graft preservation.MethodsPig liver grafts were preserved using SCS, HOPE, or SCS+M101, and the liver functions were compared during cold preservation and after orthotopic allotransplantation (OLT) in pigs.ResultsDuring preservation of the liver grafts, mitochondrial function, ATP synthesis, antioxidant capacities, and hepatocyte architecture were better preserved, and free radical production, antioxidant activities, and inflammatory mediators were lower, with HOPE or SCS+M101 than with SCS alone. However, after 1 h of preservation, liver functions with HOPE were superior to those with SCS+M101. After 6 h of preservation and OLT, blood levels of aspartate and alanine aminotransferases and lactate dehydrogenase increased with a peak effect at Day 1 post-transplant; values were similar with HOPE and SCS+M101, and were significantly lower than those in the SCS group. At Days 1 and 3, tumor necrosis factor α levels remained lower with HOPE and SCS+M101 vs. SCS. At Day 7, liver cell necrosis and inflammation were less marked in both oxygenated groups.ConclusionsWhen added to SCS, M101 effectively oxygenates liver grafts during preservation, preventing post-transplant injury; although graft performances are below those achieved with HOPE.Lay summaryWhen transported between donors and recipients, even cold-stored liver grafts need oxygen to maintain their viability. To provide them with oxygen, we added a marine worm super haemoglobin (M101) to the cold-storage solution UWCS. Using a pig liver transplant model, we revealed that livers cold stored with UWCS+M101 showed improved oxygenation compared with simple cold-storage solutions, but did not reach the oxygenation level achieved with machine perfusion.

Effects of Guanidinoacetic acid supplements on laying performance, egg quality, liver nitric oxide and energy metabolism in laying hens at the late stage of production

AbstractThe objective was to elucidate the effects of dietary supplementation with guanidinoacetic acid (GAA) on performance, egg quality and the liver antioxidant activity of laying hens. A total of 128, 72-week-old ISA Brown laying hens were randomly divided into four equal groups (32 birds), and each subgroup had eight replicates (four birds/cage). The control group (GAA0) fed the basal diet with no supplements, while the other experimental groups fed the basal diets supplemented with 0.5 (GAA1), 1.0 (GAA2) and 1.5 (GAA3) g of GAA/kg diet. The experiment lasted for 6 weeks. The addition of GAA at a rate of 1.5 g kg−1 significantly increased the hen-day egg production and egg mass as compared to the control group (P = 0.016 and 0.003, respectively). Although the egg weight was not affected (P = 0.521) by the dietary supplements, the shell ratio, shell thickness, yolk index and Haugh units increased linearly with the increase in the dietary supplements of the GAA (P = 0.036, 0.001, 0.012 and 0.004, respectively). The liver MDA levels decreased linearly with the increment in the dietary levels of the GAA (P = 0.012). Birds in the GAA2 and GAA3 showed a significantly higher liver nitric oxide level (52.50 and 54.21 mg/g, respectively) when compared with GAA0 and GAA1 groups (P = 0.029). Compared to the GAA0 group, all GAA-supplemented groups showed significantly higher liver ATP levels (P = 0.047). In conclusion, the dietary GA supplements at doses of 1.0 or 1.5 g kg−1 may improve the laying performance, antioxidant activity and the status of cellular energy metabolism in laying hens.

Microvasculopathy and soft tissue calcification in mice are governed by fetuin-A, magnesium and pyrophosphate.

Calcifications can disrupt organ function in the cardiovascular system and the kidney, and are particularly common in patients with chronic kidney disease (CKD). Fetuin-A deficient mice maintained against the genetic background DBA/2 exhibit particularly severe soft tissue calcifications, while fetuin-A deficient C57BL/6 mice remain healthy. We employed molecular genetic analysis to identify risk factors of calcification in fetuin-A deficient mice. We sought to identify pharmaceutical therapeutic targets that could be influenced by dietary of parenteral supplementation. We studied the progeny of an intercross of fetuin-A deficient DBA/2 and C57BL/6 mice to identify candidate risk genes involved in calcification. We determined that a hypomorphic mutation of the Abcc6 gene, a liver ATP transporter supplying systemic pyrophosphate, and failure to regulate the Trpm6 magnesium transporter in kidney were associated with severity of calcification. Calcification prone fetuin-A deficient mice were alternatively treated with parenteral administration of fetuin-A dietary magnesium supplementation, phosphate restriction, or by or parenteral pyrophosphate. All treatments markedly reduced soft tissue calcification, demonstrated by computed tomography, histology and tissue calcium measurement. We show that pathological ectopic calcification in fetuin-A deficient DBA/2 mice is caused by a compound deficiency of three major extracellular and systemic inhibitors of calcification, namely fetuin-A, magnesium, and pyrophosphate. All three of these are individually known to contribute to stabilize protein-mineral complexes and thus inhibit mineral precipitation from extracellular fluid. We show for the first time a compound triple deficiency that can be treated by simple dietary or parenteral supplementation. This is of special importance in patients with advanced CKD, who commonly exhibit reduced serum fetuin-A, magnesium and pyrophosphate levels.

Hepatocyte-Specific Deletion of AMPKα1 Results in Worse Outcomes in Mice Subjected to Sepsis in a Sex-Specific Manner.

Alterations in the energy homeostasis contribute to sepsis-mediated multiple organ failure. The liver plays a central role in metabolism and participates to the innate immune and inflammatory responses of sepsis. Several clinical and experimental studies have suggested that females are less susceptible to the adverse outcome of sepsis. However, underlying mechanisms of organ damage in sepsis remain largely undefined. AMP-activated protein kinase (AMPK) is an important regulator of mitochondrial quality control. The AMPK catalytic α1 isoform is abundantly expressed in the liver. Here, we determined the role of hepatocyte AMPKα1 in sepsis by using hepatocyte-specific AMPKα1 knockout mice (H-AMPKα1 KO) generated with Cre-recombinase expression under the control of the albumin promoter. Using a clinically relevant model of polymicrobial sepsis by cecal ligation and puncture (CLP), we observed that male H-AMPKα1 KO mice had higher plasma levels of tumor necrosis factor-α and interleukin-6 and exhibited a more severe liver and lung injury than male H-AMPKα1 WT mice, as evaluated by histology and neutrophil infiltration at 18 h after CLP. Plasma levels of interleukin-10 and the keratinocyte-derived chemokine were similarly elevated in both KO and WT male mice. At transmission electron microscopy analysis, male H-AMPKα1 KO mice exhibited higher liver mitochondrial damage, which was associated with a significant decrease in liver ATP levels when compared to WT mice at 18 h after sepsis. Mortality rate was significantly higher in the male H-AMPKα1 KO group (91%) when compared to WT mice (60%) at 7 days after CLP. Female H-AMPKα1 WT mice exhibited a similar degree of histological liver and lung injury, but significantly milder liver mitochondrial damage and higher autophagy when compared to male WT mice after CLP. Interestingly, H-AMPKα1 KO female mice had lower organ neutrophil infiltration, lower liver mitochondrial damage and lower levels of cytokines than WT female mice. There was no significant difference in survival rate between WT and KO mice in the female group. In conclusion, our study demonstrates that AMPKα1 is a crucial hepatoprotective enzyme during sepsis. Furthermore, our results suggest that AMPK-dependent liver metabolic functions may influence the susceptibility to multiple organ injury in a sex-dependent manner.

Liver ChREBP Protects Against Fructose-Induced Glycogenic Hepatotoxicity by Regulating L-Type Pyruvate Kinase.

Excessive fructose consumption is closely linked to the pathogenesis of metabolic disease. Carbohydrate response element-binding protein (ChREBP) is a transcription factor essential for fructose tolerance in mice. However, the functional significance of liver ChREBP in fructose metabolism remains unclear. Here, we show that liver ChREBP protects mice against fructose-induced hepatotoxicity by regulating liver glycogen metabolism and ATP homeostasis. Liver-specific ablation of ChREBP did not compromise fructose tolerance, but rather caused severe transaminitis and hepatomegaly with massive glycogen overload in mice fed a high-fructose diet, while no obvious inflammation, cell death, or fibrosis was detected in the liver. In addition, liver ATP contents were significantly decreased by ChREBP deficiency in the fed state, which was rendered more pronounced by fructose feeding. Mechanistically, liver contents of glucose-6-phosphate (G6P), an allosteric activator of glycogen synthase, were markedly increased in the absence of liver ChREBP, while fasting-induced glycogen breakdown was not compromised. Furthermore, hepatic overexpression of LPK, a ChREBP target gene in glycolysis, could effectively rescue glycogen overload and ATP reduction, as well as mitigate fructose-induced hepatotoxicity in ChREBP-deficient mice. Taken together, our findings establish a critical role of liver ChREBP in coping with hepatic fructose stress and protecting from hepatotoxicity by regulating LPK.

Adenine nucleotide-mediated regulation of hepatic PTP1B activity in mouse models of type 2 diabetes.

Plasma 5'-AMP (pAMP) is elevated in mouse models of type 2 diabetes. However, the metabolic regulatory role of adenine nucleotides in type 2 diabetes remains unclear. Adenine nucleotides and their metabolites in plasma and liver were examined by HPLC. 1H NMR-based metabolomics analysis was performed to explore the changes of metabolites in mouse models of type 2 diabetes. Na+/K+ ATPase and Na+/H+ exchanger activity were measured in response to adenine nucleotide metabolites. Human recombinant protein tyrosine phosphatase 1B (PTP1B) was used for enzyme kinetic assays. Protein binding assays were performed with microscale thermophoresis. The intracellular pH of hepatocyte AML12 cell lines was measured using the BCECF-AM method. We also analysed pAMP levels in participants with type 2 diabetes. Elevation of pAMP was a universal phenomenon in all mouse models of type 2 diabetes including db/db vs lean mice (13.9 ± 2.3μmol/l vs 3.7 ± 0.9μmol/l; p < 0.01), ob/ob vs lean mice (9.1 ± 2.0μmol/l vs 3.9 ± 1.2μmol/l; p < 0.01) and high-fat diet/streptozotocin-induced vs wild-type mice (6.6 ± 1.5μmol/l vs 4.1 ± 0.9μmol/l; p < 0.05); this elevation was required for the occurrence of hyperglycaemia in obese mice. 1H NMR-based metabolomics study following HPLC analysis revealed that the metabolite profile in wild-type mice treated with 5'-AMP was similar to that in db/db diabetic mice, especially the accumulation of a large quantity of ATP and its metabolites. The glucose-lowering drug metformin reduced the severity of hyperglycaemia both in 5'-AMP-induced wild-type mice and db/db mice. Metformin decreased the accumulation of liver ATP but not its metabolites in these hyperglycaemic mice. ATP and metformin reciprocally change cellular pH homeostasis in liver, causing opposite shifts in liver activity of PTP1B, a key negative regulator of insulin signalling. Furthermore, pAMP levels were also elevated in individuals with type 2 diabetes (45.2 ± 22.7nmol/l vs 3.1 ± 1.9nmol/l; p < 0.01). These results reveal an emerging role for adenine nucleotide in the regulation of hyperglycaemia and provide a potential therapeutic target in obesity and type 2 diabetes.

Decreased hepatic response to glucagon, adrenergic agonists, and cAMP in glycogenolysis, gluconeogenesis, and glycolysis in tumor-bearing rats.

The response to glucagon and adrenaline in cancer cachexia is poorly known. The aim of this study was to investigate the response to glucagon, adrenergic agonists (α and β) and cyclic adenosine monophosphate (cAMP) on glycogenolysis, gluconeogenesis, and glycolysis in liver perfusion of Walker-256 tumor-bearing rats with advanced cachexia. Liver ATP content was also investigated. Rats without tumor (healthy) were used as controls. Agonists α (phenylephrine) and β (isoproterenol) adrenergic, instead of adrenaline, and cAMP, the second messenger of glucagon and isoproterenol, were used in an attempt to identify mechanisms involved in the responses. Glucagon (1 nM) stimulated glycogenolysis and gluconeogenesis and inhibited glycolysis in the liver of healthy and tumor-bearing rats, but their effects were lower in tumor-bearing rats. Isoproterenol (20 µM) stimulated glycogenolysis, gluconeogenesis, and glycolysis in healthy rats and had virtually no effect in tumor-bearing rats. cAMP (9 µM) also stimulated glycogenolysis and gluconeogenesis and inhibited glycolysis in healthy rats but had practically no effect in tumor-bearing rats. Phenylephrine (2 µM) stimulated glycogenolysis and gluconeogenesis and inhibited glycolysis and these effects were also lower in tumor-bearing rats than in healthy. Liver ATP content was lower in tumor-bearing rats. In conclusion, tumor-bearing rats with advanced cachexia showed a decreased hepatic response to glucagon, adrenergic agonists (α and β), and cAMP in glycogenolysis, gluconeogenesis, and glycolysis, which may be due to a reduced rate of regulatory enzyme phosphorylation caused by the low ATP levels in the liver.

Chronic stress, energy transduction, and free-radical production in a reptile.

Stress hormones, such as corticosterone, play a crucial role in orchestrating physiological reaction patterns shaping adapted responses to stressful environments. Concepts aiming at predicting individual and population responses to environmental stress typically consider that stress hormones and their effects on metabolic rate provide appropriate proxies for the energy budget. However, uncoupling between the biochemical processes of respiration, ATP production, and free-radical production in mitochondria may play a fundamental role in the stress response and associated life histories. In this study, we aim at dissecting sub-cellular mechanisms that link these three processes by investigating both whole-organism metabolism, liver mitochondrial oxidative phosphorylation processes (O2 consumption and ATP production) and ROS emission in Zootoca vivipara individuals exposed 21days to corticosterone relative to a placebo. Corticosterone enhancement had no effect on mitochondrial activity and efficiency. In parallel, the corticosterone treatment increased liver mass and mitochondrial protein content suggesting a higher liver ATP production. We also found a negative correlation between mitochondrial ROS emission and plasma corticosterone level. These results provide a proximal explanation for enhanced survival after chronic exposure to corticosterone in this species. Importantly, none of these modifications affected resting whole-body metabolic rate. Oxygen consumption, ATP, and ROS emission were thus independently affected in responses to corticosterone increase suggesting that concepts and models aiming at linking environmental stress and individual responses may misestimate energy allocation possibilities.

Regulation of energy metabolism during social interactions in rainbow trout: a role for AMP-activated protein kinase.

Rainbow trout (Oncorhynchus mykiss) confined in pairs form social hierarchies in which subordinate fish typically experience fasting and high circulating cortisol levels, resulting in low growth rates. The present study investigated the role of AMP-activated protein kinase (AMPK) in mediating metabolic adjustments associated with social status in rainbow trout. After 3 days of social interaction, liver AMPK activity was significantly higher in subordinate than dominant or sham (fish handled in the same fashion as paired fish but held individually) trout. Elevated liver AMPK activity in subordinate fish likely reflected a significantly higher ratio of phosphorylated AMPK (phospho-AMPK) to total AMPK protein, which was accompanied by significantly higher AMPKα1 relative mRNA abundance. Liver ATP and creatine phosphate concentrations in subordinate fish also were elevated, perhaps as a result of AMPK activity. Sham fish that were fasted for 3 days exhibited effects parallel to those of subordinate fish, suggesting that low food intake was an important trigger of elevated AMPK activity in subordinate fish. Effects on white muscle appeared to be influenced by the physical activity associated with social interaction. Overall, muscle AMPK activity was significantly higher in dominant and subordinate than sham fish. The ratio of phospho-AMPK to total AMPK protein in muscle was highest in subordinate fish, while muscle AMPKα1 relative mRNA abundance was elevated by social dominance. Muscle ATP and creatine phosphate concentrations were high in dominant and subordinate fish at 6 h of interaction and decreased significantly thereafter. Collectively, the findings of the present study support a role for AMPK in mediating liver and white muscle metabolic adjustments associated with social hierarchy formation in rainbow trout.

Lack of Additive Effects of Resveratrol and Energy Restriction in the Treatment of Hepatic Steatosis in Rats.

The aims of the present study were to analyze the effect of resveratrol on liver steatosis in obese rats, to compare the effects induced by resveratrol and energy restriction and to research potential additive effects. Rats were initially fed a high-fat high-sucrose diet for six weeks and then allocated in four experimental groups fed a standard diet: a control group, a resveratrol-treated group, an energy restricted group and a group submitted to energy restriction and treated with resveratrol. We measured liver triacylglycerols, transaminases, FAS, MTP, CPT1a, CS, COX, SDH and ATP synthase activities, FATP2/FATP5, DGAT2, PPARα, SIRT1, UCP2 protein expressions, ACC and AMPK phosphorylation and PGC1α deacetylation. Resveratrol reduced triacylglycerols compared with the controls, although this reduction was lower than that induced by energy restriction. The mechanisms of action were different. Both decreased protein expression of fatty acid transporters, thus suggesting reduced fatty acid uptake from blood stream and liver triacylglycerol delivery, but only energy restriction reduced the assembly. These results show that resveratrol is useful for liver steatosis treatment within a balanced diet, although its effectiveness is lower than that of energy restriction. However, resveratrol is unable to increase the reduction in triacylglycerol content induced by energy restriction.