RESUMO
l-carnitine is a key molecule in both mitochondrial and peroxisomal lipid metabolisms. l-carnitine is biosynthesized from gamma-butyrobetaine by a reaction catalyzed by the gamma-butyrobetaine hydroxylase (Bbox1). The aim of this work was to identify molecular mechanisms involved in the regulation of l-carnitine biosynthesis and availability. Using 3' RACE, we identified four alternatively polyadenylated Bbox1 mRNAs in rat liver. We utilized a combination of in vitro experiments using hybrid constructs containing the Bbox1 3' UTR and in vivo experiments on rat liver mRNAs to reveal specificities in the different Bbox1 mRNA isoforms, especially in terms of polyadenylation efficiency, mRNA stability and translation efficiency. This complex maturation process of the Bbox1 mRNAs in the liver was studied on rats fed a high-fat diet. High-fat diet selectively increased the level of three Bbox1 mRNA isoforms in rat liver and the alternative use of polyadenylation sites contributed to the global increase in Bbox1 enzymatic activity and l-carnitine levels. Our results show that the maturation of Bbox1 mRNAs is nutritionally regulated in the liver through a selective polyadenylation process to adjust l-carnitine biosynthesis to the energy supply.
Assuntos
Carnitina/biossíntese , Gorduras na Dieta/administração & dosagem , RNA Mensageiro/genética , Animais , Sequência de Bases , Linhagem Celular , Primers do DNA , Fígado/metabolismo , Masculino , Dados de Sequência Molecular , Ratos , Ratos WistarRESUMO
This editorial explores the intricate relationship between microplastics (MPs) and gut microbiota, emphasizing the complexity and environmental health implications. The gut microbiota, a crucial component of gastrointestinal health, is examined in the context of potential microbial degradation of MPs. Furthermore, dysbiosis induced by MPs emerges as a consensus, disrupting the balance of gut microbiota and decreasing diversity. The mechanisms triggering dysbiosis, including physical interactions and chemical composition, are under investigation. Ongoing research addresses the consequences of MPs on immune fun-ction, nutrient metabolism, and overall host health. The bidirectional relationship between MPs and gut microbiota has significant implications for environmental and human health. Despite uncertainties, MPs negatively impact gut microbiota and health. Further research is essential to unravel the complex interactions and assess the long-term consequences of MPs on both environmental and human well-being.
Assuntos
Disbiose , Microbioma Gastrointestinal , Microplásticos , Microplásticos/efeitos adversos , Humanos , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/fisiologia , Saúde Ambiental , Exposição Ambiental/efeitos adversos , AnimaisRESUMO
The peroxisomal beta oxidation of very long chain fatty acids (VLCFA) leads to the formation of medium chain acyl-CoAs such as octanoyl-CoA. Today, it seems clear that the exit of shortened fatty acids produced by the peroxisomal beta oxidation requires their conversion into acyl-carnitine and the presence of the carnitine octanoyltransferase (CROT). Here, we describe the consequences of an overexpression and a knock down of the CROT gene in terms of mitochondrial and peroxisomal fatty acids metabolism in a model of hepatic cells. Our experiments showed that an increase in CROT activity induced a decrease in MCFA and VLCFA levels in the cell. These changes are accompanied by an increase in the level of mRNA encoding enzymes of the peroxisomal beta oxidation. In the same time, we did not observe any change in mitochondrial function. Conversely, a decrease in CROT activity had the opposite effect. These results suggest that CROT activity, by controlling the peroxisomal amount of medium chain acyls, may control the peroxisomal oxidative pathway.
Assuntos
Carnitina Aciltransferases/fisiologia , Ácidos Graxos/metabolismo , Peroxissomos/enzimologia , Carnitina Aciltransferases/genética , Técnicas de Silenciamento de Genes , Células HEK293 , Células Hep G2 , Humanos , Oxirredução , RNA Interferente Pequeno/genéticaRESUMO
Unbalanced energy partitioning participates in the rise of obesity, a major public health concern in many countries. Increasing basal energy expenditure has been proposed as a strategy to fight obesity yet raises efficiency and safety concerns. Here, we show that mice deficient for a muscle-specific enzyme of very-long-chain fatty acid synthesis display increased basal energy expenditure and protection against high-fat diet-induced obesity. Mechanistically, muscle-specific modulation of the very-long-chain fatty acid pathway was associated with a reduced content of the inner mitochondrial membrane phospholipid cardiolipin and a blunted coupling efficiency between the respiratory chain and adenosine 5'-triphosphate (ATP) synthase, which was restored by cardiolipin enrichment. Our study reveals that selective increase of lipid oxidative capacities in skeletal muscle, through the cardiolipin-dependent lowering of mitochondrial ATP production, provides an effective option against obesity at the whole-body level.
RESUMO
Patients with autism spectrum disorders (ASD) present deficits in social interactions and communication, they also show limited and stereotypical patterns of behaviors and interests. The pathophysiological bases of ASD have not been defined yet. Many factors seem to be involved in the onset of this disorder. These include genetic and environmental factors, but autism is not linked to a single origin, only. Autism onset can be connected with various factors such as metabolic disorders: including carnitine deficiency. Carnitine is a derivative of two amino acid lysine and methionine. Carnitine is a cofactor for a large family of enzymes: the carnitine acyltransferases. Through their action these enzymes (and L-carnitine) are involved in energy production and metabolic homeostasis. Some people with autism (less than 20%) seem to have L-carnitine metabolism disorders and for these patients, a dietary supplementation with L-carnitine is beneficial. This review summarizes the available information on this topic.
RESUMO
L-carnitine is an essential cofactor for the transport of fatty acids across the mitochondrial membranes. L-carnitine can be provided by food products or biosynthesized in the liver. After intestinal absorption or hepatic biosynthesis, L-carnitine is transferred to organs whose metabolism is dependent upon fatty acid oxidation, such as the skeletal muscle and the heart. The intracellular transport of L-carnitine into the cell requires specific transporters and today, several of these have been characterized. Most of them belong to the solute carrier family. Heart is one of the major target for carnitine transport and use, however basic properties of carnitine uptake by heart cells have never been studied. In this paper, the transport of L-carnitine by rat heart explants has been examined and the kinetic properties of this transport determined and compared to data obtained in skeletal muscle explants. As in muscle, L-carnitine uptake by heart cells was shown to be dependent on sodium and was inhibited by L-carnitine analogues. Molecules known to interact with the skeletal muscle L-carnitine transport were studied in the heart. While trimethyl hydrazinium propionate (THP) was shown to fully inhibit the L-carnitine uptake by muscle cells, it remained inefficient in inhibiting the L-carnitine uptake by heart cells. On the other hand, compounds such as verapamil and AZT were both able to inhibit both the skeletal muscle and the cardiac uptake of L-carnitine. These data suggested that the muscle and heart systems for L-carnitine uptake exhibited different systems of regulation and these results have to be taken in consideration while administrating those compounds that can alter l-carnitine uptake in the muscle and the heart and can lead to damage to these tissues.
Assuntos
Carnitina/metabolismo , Miócitos Cardíacos/metabolismo , Animais , Masculino , Ratos , Ratos Wistar , Sódio/metabolismoRESUMO
In mammals, during the aging process, an atrophy of the muscle fibers, an increase in body fat mass, and a decrease in skeletal muscle oxidative capacities occur. Compounds and activities that interact with lipid oxidative metabolism may be useful in limiting damages that occur in aging muscle. In this study, we evaluated the effect of L-carnitine and physical exercise on several parameters related to muscle physiology. We described that supplementing old rats with L-carnitine at 30 mg/kg body weight for 12 weeks (a) allowed the restoration of L-carnitine level in muscle cells, (b) restored muscle oxidative activity in the soleus, and (c) induced positive changes in body composition: a decrease in abdominal fat mass and an increase in muscle capabilities without any change in food intake. Moderate physical exercise was also effective in (a) limiting fat mass gain and (b) inducing an increase in the capacities of the soleus to oxidize fatty acids.
Assuntos
Envelhecimento/efeitos dos fármacos , Envelhecimento/fisiologia , Carnitina/farmacologia , Mitocôndrias/metabolismo , Músculo Esquelético/fisiologia , Condicionamento Físico Animal , Animais , Masculino , Músculo Esquelético/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Estatísticas não ParamétricasRESUMO
Extracellular ATP regulates cell proliferation, muscle contraction and myoblast differentiation. ATP present in the muscle interstitium can be released from contracting skeletal muscle cells. L-Carnitine is a key element in muscle cell metabolism, as it serves as a carrier for fatty acid through mitochondrial membranes, controlling oxidation and energy production. Treatment of C2C12 cells with 1 mmol/l of ATP induced a marked increase in L-carnitine uptake that was associated with an increase in L-carnitine content in these cells. These effects were found to be dependent on the density of the cultured cells and on the dose of ATP. The use of specific inhibitors of P2X and P2Y receptors abolished the effect of ATP on L-carnitine metabolism. As ATP can be released from stressed or exercising cells, it can be hypothesized that ATP acts as a messenger in the muscle. ATP will be released to recruit the next cells and increase their metabolism.
Assuntos
Trifosfato de Adenosina/metabolismo , Carnitina/metabolismo , Receptores Purinérgicos P2/metabolismo , Complexo Vitamínico B/metabolismo , Trifosfato de Adenosina/administração & dosagem , Animais , Transporte Biológico , Linhagem Celular , Relação Dose-Resposta a Droga , Camundongos , Mioblastos/metabolismo , Antagonistas do Receptor Purinérgico P2RESUMO
Human adults store around 20g of l-carnitine. In the human body, l-carnitine is not metabolized but excreted through the kidney. Lost l-carnitine has to be replenished either by a biosynthetic mechanism or by the consumption of foods containing l-carnitine. Today, there is no "official" recommended daily allowance for l-carnitine but the daily need for l-carnitine intake has been estimated in the wide range of 2-12µmol/day/kg body weight for an adult human. In this study we evaluated the effect of freezing and of different cooking methods on the l-carnitine content of red meat and fish. l-carnitine was abundantly present in all beef products analyzed. The amounts in the various cuts were similar and our data showed that freezing or cooking did not modify l-carnitine content. Salmon contained about 12 times less l-carnitine than beef but except in smoked salmon, cooking or freezing did not alter l-carnitine content. This study confirms the important role that meet products play for providing adequate amount of l-carnitine to the human body.
RESUMO
Gamma-butyrobetaine hydroxylase (BBOX1) is the enzyme responsible for the biosynthesis of l-carnitine, a key molecule of fatty acid metabolism. This cytosolic dimeric protein belongs to the dioxygenase family. In human, enzyme activity has been detected in kidney, liver and brain. The human gene encoding gamma-butyrobetaine hydroxylase is located on chromosome 11. Although the protein structure and activity have been extensively described, little information is available concerning BBOX1 structure and expression. In this study, the organization of the human gene was determined. The structure and functions of the 5'- and 3'-untranslated regions of the human BBOX1 mRNA were characterized in kidney, liver and brain. Our experiments revealed that the transcription initiation of the human BBOX1 gene might occur at 3 different exons, and that the expression level of each type of transcript is organ-specific. We showed that the use of 3 different promoters is responsible for the 5'-end heterogeneity. Investigations on BBOX1 mRNA maturation highlighted an alternative polyadenylation mechanism that generates two 3'-untranslated regions differing by their length. This alternative polyadenylation exhibited a tissue specificity.
Assuntos
gama-Butirobetaína Dioxigenase/genética , Regiões 3' não Traduzidas , Regiões 5' não Traduzidas , Processamento Alternativo , Sequência de Bases , Encéfalo/enzimologia , DNA Complementar/genética , Éxons , Etiquetas de Sequências Expressas , Expressão Gênica , Humanos , Rim/enzimologia , Fígado/enzimologia , Dados de Sequência Molecular , Regiões Promotoras Genéticas , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Tecidual , Transcrição Gênica , gama-Butirobetaína Dioxigenase/química , gama-Butirobetaína Dioxigenase/metabolismoRESUMO
AIM: To identify and characterize the protective effect that L-carnitine exerted against an oxidative stress in C2C12 cells. METHODS: Myoblastic C2C12 cells were treated with menadione, a vitamin K analog that engenders oxidative stress, and the protective effect of L-carnitine (a nutrient involved in fatty acid metabolism and the control of the oxidative process), was assessed by monitoring various parameters related to the oxidative stress, autophagy and cell death. RESULTS: Associated with its physiological function, a muscle cell metabolism is highly dependent on oxygen and may produce reactive oxygen species (ROS), especially under pathological conditions. High levels of ROS are known to induce injuries in cell structure as they interact at many levels in cell function. In C2C12 cells, a treatment with menadione induced a loss of transmembrane mitochondrial potential, an increase in mitochondrial production of ROS; it also induces autophagy and was able to provoke cell death. Pre-treatment of the cells with L-carnitine reduced ROS production, diminished autophagy and protected C2C12 cells against menadione-induced deleterious effects. CONCLUSION: In conclusion, L-carnitine limits the oxidative stress in these cells and prevents cell death.
RESUMO
Mitochondria and peroxisomes are small ubiquitous organelles. They both play major roles in cell metabolism, especially in terms of fatty acid metabolism, reactive oxygen species (ROS) production, and ROS scavenging, and it is now clear that they metabolically interact with each other. These two organelles share some properties, such as great plasticity and high potency to adapt their form and number according to cell requirements. Their functions are connected, and any alteration in the function of mitochondria may induce changes in peroxisomal physiology. The objective of this paper was to highlight the interconnection and the crosstalk existing between mitochondria and peroxisomes. Special emphasis was placed on the best known connections between these organelles: origin, structure, and metabolic interconnections.
RESUMO
The reduced diameter of skeletal myofibres is a hallmark of several congenital myopathies, yet the underlying cellular and molecular mechanisms remain elusive. In this study, we investigate the role of HACD1/PTPLA, which is involved in the elongation of the very long chain fatty acids, in muscle fibre formation. In humans and dogs, HACD1 deficiency leads to a congenital myopathy with fibre size disproportion associated with a generalized muscle weakness. Through analysis of HACD1-deficient Labradors, Hacd1-knockout mice, and Hacd1-deficient myoblasts, we provide evidence that HACD1 promotes myoblast fusion during muscle development and regeneration. We further demonstrate that in normal differentiating myoblasts, expression of the catalytically active HACD1 isoform, which is encoded by a muscle-enriched splice variant, yields decreased lysophosphatidylcholine content, a potent inhibitor of myoblast fusion, and increased concentrations of ≥ C18 and monounsaturated fatty acids of phospholipids. These lipid modifications correlate with a reduction in plasma membrane rigidity. In conclusion, we propose that fusion impairment constitutes a novel, non-exclusive pathological mechanism operating in congenital myopathies and reveal that HACD1 is a key regulator of a lipid-dependent muscle fibre growth mechanism.
Assuntos
Membrana Celular/metabolismo , Desenvolvimento Muscular/fisiologia , Mioblastos/citologia , Proteínas Tirosina Fosfatases/metabolismo , Animais , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Linhagem Celular , Membrana Celular/genética , Cães , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Desenvolvimento Muscular/genética , Músculo Esquelético/metabolismo , Mioblastos/metabolismo , Proteínas Tirosina Fosfatases/genéticaRESUMO
L-Carnitine is a key molecule in the transfer of fatty acid across mitochondrial membranes. Bioavailable L-carnitine is either provided by an endogeneous biosynthesis or after intestinal absorption of dietary items containing L-carnitine. After intestinal absorption or hepatic biosynthesis, L-carnitine is transferred to organs whose metabolism is dependent upon fatty acid oxidation, such as skeletal muscle. To cross the muscle plasma membrane, there are several transporters involved. Among those transporters, OCTN2 is actually the only one to have been clearly characterized. Zidovudine is a commonly used inhibitor of human immunodeficiency virus (HIV) replication. Zidovudine has many side effects, including induction of myopathy characterized by a metabolic mitochondria dysfunction and a diminution of the muscle L-carnitine content. In this study, we described the characteristics of L-carnitine transport in C2C12 cells. We also demonstrated that zidovudine inhibited the L-carnitine transporter. This inhibition led to a significant reduction of the muscle cell growth. In C2C12 cells, the supplementation of L-carnitine prevented the effects of zidovudine and restored the normal cell growth.
Assuntos
Antimetabólitos/farmacologia , Carnitina/metabolismo , Mioblastos/efeitos dos fármacos , Zidovudina/farmacologia , Animais , Antimetabólitos/efeitos adversos , Transporte Biológico , Divisão Celular/efeitos dos fármacos , Interações Medicamentosas , Cinética , Camundongos , Especificidade por Substrato , Zidovudina/efeitos adversosRESUMO
In Alzheimer's disease (AD) and dementia of the Alzheimer's type (DAT), the role played by peroxisomes is not well known. Peroxisomes are present in all eukaryotic cells, with the exception of erythrocytes. They are involved in the ß-oxidation process of long-chain fatty acids, very-long-chain fatty acids, and branched-chain fatty acids. They participate in the α-oxidation of phytanic acid, the biosynthesis of bile acids, and the breakdown of eicosanoids. Peroxisomes are also involved in the synthesis of specific fatty acids such as docosahexaenoic acid (DHA), which is essential for the brain and retina, and plasmalogens (PLGN), which play crucial roles in neural cells and are essential components of myelin. Several studies conducted in animal models and in humans provided evidence for a role of DHA in preventing brain degeneration. Significantly lower levels of PLGN were observed in patients with severe dementia. Moreover, a decreased activity of carnitine acetyltransferase, an enzyme present in peroxisome (but also detected in mitochondria, endoplasmic reticulum, and nucleus), was reported in AD patients. We give an overview of the potential role of peroxisomes, especially in the part played by DHA, PLGN, carnitine, and carnitine-dependent peroxisomal enzymes, on the development of AD and DAT. The potential of developing novel therapies targeted on peroxisomal metabolism to prevent cognitive decline and other age-related neurological disorders is discussed.
Assuntos
Doença de Alzheimer/patologia , Encéfalo/patologia , Neurônios/patologia , Neurônios/ultraestrutura , Peroxissomos/metabolismo , Animais , Humanos , Mitocôndrias/metabolismo , Modelos Biológicos , Oxirredução , Peroxissomos/ultraestruturaRESUMO
Fatty acids are known to serve as energetic substrates, key components of membrane lipids, and as substrates for the synthesis of signaling molecules and complex lipids. They are also known to be ligands either of membrane receptors involved in cell signaling or of nuclear receptors mediating gene regulation. Accumulation of fatty acids due to altered metabolism and/or unbalanced diet has been described to be toxic for several tissues, especially liver. In numerous cell types, cell death, cytokine secretion and activation of inflammatory processes appear to be a consequence of fatty acid accumulation. This review presents the different classes of fatty acids known to trigger toxic effects and inflammation, the cellular and subcellular targets of these fatty acids in the context of non-alcoholic fatty liver disease (NAFLD), and the mechanisms by which these effects are mediated.
Assuntos
Ácidos Graxos/metabolismo , Fígado Gorduroso/metabolismo , Inflamação/metabolismo , Animais , Hepatócitos/metabolismo , Humanos , Hepatopatia Gordurosa não Alcoólica , Estresse OxidativoRESUMO
Duchenne muscular dystrophy (DMD) arises as a consequence of mutations in the dystrophin gene. Dystrophin is a membrane-spanning protein that connects the cytoskeleton and the basal lamina. The most distinctive features of DMD are a progressive muscular dystrophy, a myofiber degeneration with fibrosis and metabolic alterations such as fatty infiltration, however, little is known on lipid metabolism changes arising in Duchenne patient cells. Our goal was to identify metabolic changes occurring in Duchenne patient cells especially in terms of L-carnitine homeostasis, fatty acid metabolism both at the mitochondrial and peroxisomal level and the consequences on the membrane structure and function. In this paper, we compared the structural and functional characteristics of DMD patient and control cells. Using radiolabeled L-carnitine, we found, in patient muscle cells, a marked decrease in the uptake and the intracellular level of L-carnitine. Associated with this change, a decrease in the mitochondrial metabolism can be seen from the analysis of mRNA encoding for mitochondrial proteins. Probably, associated with these changes in fatty acid metabolism, alterations in the lipid composition of the cells were identified: with an increase in poly unsaturated fatty acids and a decrease in medium chain fatty acids, mono unsaturated fatty acids and in cholesterol contents. Functionally, the membrane of cells lacking dystrophin appeared to be less fluid, as determined at 37°C by fluorescence anisotropy. These changes may, at least in part, be responsible for changes in the phospholipids and cholesterol profile in cell membranes and ultimately may reduce the fluidity of the membrane. A supplementation with L-carnitine partly restored the fatty acid profile by increasing saturated fatty acid content and decreasing the amounts of MUFA, PUFA, VLCFA. L-carnitine supplementation also restored muscle membrane fluidity. This suggests that regulating lipid metabolism in DMD cells may improve the function of cells lacking dystrophin.
Assuntos
Carnitina/metabolismo , Membrana Celular/metabolismo , Metabolismo dos Lipídeos , Distrofia Muscular de Duchenne/metabolismo , Adolescente , Membrana Celular/química , Ácidos Graxos/metabolismo , Humanos , Masculino , Mitocôndrias/genética , Mitocôndrias/metabolismo , Células Musculares/metabolismo , Distrofia Muscular de Duchenne/genética , Fosfolipídeos/metabolismoRESUMO
L-Carnitine plays an important role in skeletal muscle bioenergetics, and its bioavailability and thus its import may be crucial for muscle function. We studied the effect of thyroid hormone, insulin, and iron overload, hormones and nutrients known to alter muscle metabolism, on L-carnitine import into C2C12 cells. We report here L-carnitine uptake is increased by thyroid hormones and decreased by iron. Insulin was found to be ineffective in altering the L-carnitine uptake.
Assuntos
Carnitina/metabolismo , Hormônios/fisiologia , Músculo Esquelético/metabolismo , Fenômenos Fisiológicos da Nutrição/fisiologia , Northern Blotting , Linhagem Celular , Humanos , Insulina/farmacologia , Compostos de Ferro/farmacologia , Sobrecarga de Ferro/metabolismo , Músculo Esquelético/citologia , Proteínas de Transporte de Cátions Orgânicos/biossíntese , Proteínas de Transporte de Cátions Orgânicos/genética , RNA Mensageiro/biossíntese , Membro 5 da Família 22 de Carreadores de Soluto , Tri-Iodotironina/farmacologiaRESUMO
Experimental evidence indicates that n-3 fatty acids, especially the long-chain polyunsaturated fatty acids, unlike n-6 fatty acids could prevent cancer development. This survey shows that fatty acids could act through several mechanisms including the production of reactive oxygen species, the modulation of gene expression and signal transduction pathways, or the eicosanoid biosynthesis. Human genetics has underlined several polymorphisms in genes identified as possible targets of fatty acids which suggests that the link between nutritional intake and cancer prevention, especially the eventual anti-carcinogenic effects of n-3 fatty acids, depends on the genetic background. Further studies are needed to evaluate the effects of fatty acids on angiogenesis which represents a marker of a poor prognosis in cancer. Finally, the use of genomic technologies combined with nutritional strategies could provide a more understanding of the effects of n-3 fatty acid intake on cancer prevention.