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1.
Am J Physiol Heart Circ Physiol ; 318(4): H787-H800, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32056460

RESUMO

Despite advances in both medical and surgical therapies, individuals with single ventricle heart disease (SV) remain at high risk for the development of heart failure (HF). However, the molecular mechanisms underlying remodeling and eventual HF in patients with SV are poorly characterized. Cardiolipin (CL), an inner mitochondrial membrane phospholipid, is critical for proper mitochondrial function, and abnormalities in CL content and composition are known in various cardiovascular disease etiologies. The purpose of this study was to investigate myocardial CL content and composition in failing and nonfailing single right ventricle (RV) samples compared with normal control RV samples, to assess mRNA expression of CL biosynthetic and remodeling enzymes, and to quantitate relative mitochondrial copy number. A cross-sectional analysis of RV myocardial tissue from 22 failing SV (SVHF), 9 nonfailing SV (SVNF), and 10 biventricular control samples (BVNF) was performed. Expression of enzymes involved in CL biosynthesis and remodeling were analyzed using RT-qPCR and relative mitochondrial DNA copy number determined by qPCR. Normal phase high-pressure liquid chromatography coupled to electrospray ionization mass spectrometry was used to quantitate total and specific CL species. While mitochondrial copy number was not significantly different between groups, total CL content was significantly lower in SVHF myocardium compared with BVNF controls. Despite having lower total CL content however, the relative percentage of the major tetralinoleoyl CL species is preserved in SVHF samples relative to BVNF controls. Correspondingly, expression of enzymes involved in CL biosynthesis and remodeling were upregulated in SVHF samples when compared with both SVNF samples and BVNF controls.NEW & NOTEWORTHY The mechanisms underlying heart failure in the single ventricle (SV) congenital heart disease population are largely unknown. In this study we identify alterations in cardiac cardiolipin metabolism, composition, and content in children with SV heart disease. These findings suggest that cardiolipin could be a novel therapeutic target in this unique population of patients.


Assuntos
Cardiolipinas/biossíntese , Coração Univentricular/metabolismo , Cardiolipinas/genética , Criança , Pré-Escolar , DNA Mitocondrial/genética , Feminino , Ventrículos do Coração/anormalidades , Ventrículos do Coração/metabolismo , Ventrículos do Coração/patologia , Humanos , Masculino , Mitocôndrias Cardíacas/enzimologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Coração Univentricular/genética , Remodelação Ventricular
2.
J Chromatogr A ; 1619: 460918, 2020 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-32008819

RESUMO

The anionic phospholipid class of cardiolipins (CL) is increasingly attracting scientific attention in the recent years. CL can be found as a functional component of mitochondrial membranes in almost all living organisms. Changes in the CL composition are favored by oxidative stress. Based on this finding, the investigation of CL and their oxidation products in relation to various disease patterns, including neurodegenerative ones, is moving into the focus of current research. The analysis of this diverse lipid class is still challenging and requires sensitive and selective methods. In this work, we demonstrate an online two-dimensional liquid chromatography (2D-LC) approach by means of a heart-cut setup. In the first dimension, a fast hydrophilic interaction liquid chromatography (HILIC) method was developed for the separation of CL and their oxidation products from other phospholipid classes, but more important from nonpolar lipid classes, such as triacylglycerol and cholesterol. Those classes can negatively affect the electrospray ionization and also the chromatography. For the heart-cut approach, the CL fraction was selectively transferred to a loop using a six-port valve followed by the transfer to a reversed phase (RP) column in second dimension. On the RP column, the transferred CL fraction including the oxidation products were separated according to the hydrophobicity of acyl chain moieties. Matrix effects were significantly reduced compared to the one-dimensional LC-MS method. In addition, the total separation time had not to be prolonged by shifting the equilibration step of the RP column parallel to the separation in first dimension. The heart-cut LC-LC approach was applied to artificially oxidized lipid extracts of bovine heart and yeast by means of Fenton reaction. In summary, 42 species have been identified by high resolution mass spectrometry and database matching. 31 species thereof have been further characterized by MS/MS experiments.


Assuntos
Cardiolipinas/análise , Técnicas de Química Analítica/métodos , Cromatografia Líquida , Espectrometria de Massas em Tandem , Animais , Bovinos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Oxirredução , Fosfolipídeos/análise
3.
Life Sci ; 245: 117352, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32006527

RESUMO

AIMS: The depot-specific differences in lipidome of visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) reflect heterogeneity of white adipose tissue (WAT), which plays a central role in its distinct response to outside stimuli. However, the detailed lipidome of depot-specific WAT is largely unknown, especially the minor constitutes including phospholipid and sphingolipid. MATERIALS AND METHODS: To investigate this field, we applied a high-coverage targeted lipidomics approach of VAT and SAT in male C57BL/6J mice to compare the basal level of their lipid profiles. Applying microarray and quantitative real-time polymerase chain reaction, we analyzed the transcriptome of twodepot-specific WAT and verified the differences in individual genes. KEY FINDINGS: In total, 342 lipid species from 19 lipid classes were identified. Our results showed the composition of TAG and FFA were different in length of chain and saturation. Interestingly, low abundance phospholipid, sphingolipid and cardiolipin were significantly higher in SAT. Lipid correlation network analysis vindicated that TAG and phospholipid formed distinct subnet and had more connections with other lipid species. Enriched ontology analysis of gene screened from LIPID MAPS and microarray suggested the differences were mainly involved in lipid metabolism, insulin resistance and inflammatory response. SIGNIFICANCE: Our comprehensive lipidomics and transcriptomics analyses revealed differences in lipid composition and lipid metabolism of two depot-specific WAT, which would offer new insights into the investigation of heterogeneity of visceral and subcutaneous white adipose tissue.


Assuntos
Tecido Adiposo Branco/metabolismo , Gordura Intra-Abdominal/metabolismo , Lipidômica , Gordura Subcutânea/metabolismo , Transcriptoma , Tecido Adiposo Branco/química , Animais , Cardiolipinas/análise , Cardiolipinas/metabolismo , Ceramidas/análise , Ceramidas/metabolismo , Ácidos Graxos/análise , Ácidos Graxos/metabolismo , Glicerídeos/análise , Glicerídeos/metabolismo , Gordura Intra-Abdominal/química , Metabolismo dos Lipídeos , Lipídeos/análise , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fosfolipídeos/análise , Fosfolipídeos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Gordura Subcutânea/química
4.
Nat Commun ; 11(1): 1103, 2020 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-32107376

RESUMO

Lipid-protein complexes are the basis of pulmonary surfactants covering the respiratory surface and mediating gas exchange in lungs. Cardiolipin is a mitochondrial lipid overexpressed in mammalian lungs infected by bacterial pneumonia. In addition, increased oxygen supply (hyperoxia) is a pathological factor also critical in bacterial pneumonia. In this paper we fabricate a micrometer-size graphene-based sensor to measure oxygen permeation through pulmonary membranes. Combining oxygen sensing, X-ray scattering, and Atomic Force Microscopy, we show that mammalian pulmonary membranes suffer a structural transformation induced by cardiolipin. We observe that cardiolipin promotes the formation of periodic protein-free inter-membrane contacts with rhombohedral symmetry. Membrane contacts, or stalks, promote a significant increase in oxygen gas permeation which may bear significance for alveoli gas exchange imbalance in pneumonia.


Assuntos
Cardiolipinas/metabolismo , Grafite/química , Bicamadas Lipídicas/metabolismo , Oxigênio/metabolismo , Alvéolos Pulmonares/metabolismo , Animais , Permeabilidade da Membrana Celular/fisiologia , Humanos , Microscopia de Força Atômica/instrumentação , Microscopia Confocal/instrumentação , Microtecnologia/instrumentação , Pneumonia Bacteriana/fisiopatologia , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/ultraestrutura , Troca Gasosa Pulmonar/fisiologia , Espalhamento a Baixo Ângulo , Transistores Eletrônicos , Difração de Raios X/instrumentação
5.
FASEB J ; 34(1): 1465-1480, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31914590

RESUMO

Cardiolipin (CL) is a hallmark phospholipid of mitochondria and plays a significant role in maintaining the mitochondrial structure and functions. Despite the physiological importance of CL, mutant organisms, yeast, Arabidopsis, C elegans, and Drosophila, which lack CL synthase (Crls1) gene and consequently are deprived of CL, are viable. Here we report conditional Crls1-deficient mice using targeted insertion of loxP sequences flanking the functional domain of CRLS1 enzyme. Homozygous null mutant mice exhibited early embryonic lethality at the peri-implantation stage. We generated neuron-specific Crls1 knockout (cKO) mice by crossing with Camk2α-Cre mice. Neuronal loss and gliosis were gradually manifested in the forebrains, where CL levels were significantly decreased. In the surviving neurons, malformed mitochondria with bubble-like or onion-like inner membrane structures were observed. We showed decreased supercomplex assembly and reduced enzymatic activities of electron transport chain complexes in the forebrain of cKO mice, resulting in affected mitochondrial calcium dynamics, a slower rate of Ca2+ uptake and a smaller calcium retention capacity. These observations clearly demonstrate indispensable roles of CL as well as of Crls1 gene in mammals.


Assuntos
Sinalização do Cálcio , Cardiolipinas/metabolismo , Embrião de Mamíferos/metabolismo , Mitocôndrias/metabolismo , Neurônios/metabolismo , Prosencéfalo/embriologia , Animais , Cálcio/metabolismo , Cardiolipinas/genética , Embrião de Mamíferos/patologia , Camundongos , Camundongos Knockout , Mitocôndrias/genética , Mitocôndrias/patologia , Neurônios/patologia , Prosencéfalo/patologia , Transferases (Outros Grupos de Fosfato Substituídos)/deficiência , Transferases (Outros Grupos de Fosfato Substituídos)/metabolismo
6.
Artigo em Inglês | MEDLINE | ID: mdl-31672571

RESUMO

Previous studies demonstrated that loss of CL in the yeast mutant crd1Δ leads to perturbation of mitochondrial iron­sulfur (FeS) cluster biogenesis, resulting in decreased activity of mitochondrial and cytosolic Fe-S-requiring enzymes, including aconitase and sulfite reductase. In the current study, we show that crd1Δ cells exhibit decreased levels of glutamate and cysteine and are deficient in the essential antioxidant, glutathione, a tripeptide of glutamate, cysteine, and glycine. Glutathione is the most abundant non-protein thiol essential for maintaining intracellular redox potential in almost all eukaryotes, including yeast. Consistent with glutathione deficiency, the growth defect of crd1Δ cells at elevated temperature was rescued by supplementation of glutathione or glutamate and cysteine. Sensitivity to the oxidants iron (FeSO4) and hydrogen peroxide (H2O2), was rescued by supplementation of glutathione. The decreased intracellular glutathione concentration in crd1Δ was restored by supplementation of glutamate and cysteine, but not by overexpressing YAP1, an activator of expression of glutathione biosynthetic enzymes. These findings show for the first time that CL plays a critical role in regulating intracellular glutathione metabolism.


Assuntos
Cardiolipinas/metabolismo , Glutationa/biossíntese , Mitocôndrias/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Cisteína/metabolismo , Compostos Ferrosos/metabolismo , Ácido Glutâmico/metabolismo , Peróxido de Hidrogênio/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Estresse Oxidativo , Saccharomyces cerevisiae , Proteínas de Saccharomyces cerevisiae/genética , Transferases (Outros Grupos de Fosfato Substituídos)/genética , Transferases (Outros Grupos de Fosfato Substituídos)/metabolismo
7.
Biochim Biophys Acta Biomembr ; 1862(2): 183064, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31521630

RESUMO

Misfolding and aggregate formation by the tau protein has been closely related with neurotoxicity in a large group of human neurodegenerative disorders, which includes Alzheimer's disease. Here, we investigate the membrane-active properties of tau oligomers on mitochondrial membranes, using minimalist in vitro model systems. Thus, exposure of isolated mitochondria to oligomeric tau evoked a disruption of mitochondrial membrane integrity, as evidenced by a combination of organelle swelling, efflux of cytochrome c and loss of the mitochondrial membrane potential. Tau-induced mitochondrial dysfunction occurred independently of the mitochondrial permeability transition (mPT) pore complex. Notably, mitochondria were rescued by pre-incubation with 10-N-nonyl acridine orange (NAO), a molecule that specifically binds cardiolipin (CL), the signature phospholipid of mitochondrial membranes. Additionally, NAO prevented direct binding of tau oligomers to isolated mitochondria. At the same time, tau proteins exhibited high affinity to CL-enriched membranes, whilst permeabilisation of lipid vesicles also strongly correlated with CL content. Intriguingly, using single-channel electrophysiology, we could demonstrate the formation of non-selective ion-conducting tau nanopores exhibiting multilevel conductances in mito-mimetic bilayers. Taken together, the data presented here advances a scenario in which toxic cytosolic entities of tau protein would target mitochondrial organelles by associating with their CL-rich membrane domains, leading to membrane poration and compromised mitochondrial structural integrity.


Assuntos
Cardiolipinas/metabolismo , Membranas Mitocondriais/efeitos dos fármacos , Proteínas tau/farmacologia , Humanos , Membranas Mitocondriais/metabolismo , Nanoporos , Permeabilidade/efeitos dos fármacos , Ligação Proteica , Multimerização Proteica
8.
Gene ; 726: 144148, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-31647997

RESUMO

Tafazzin, which is encoded by the TAZ gene, catalyzes transacylation to form mature cardiolipin and shows preference for the transfer of a linoleic acid (LA) group from phosphatidylcholine (PC) to monolysocardiolipin (MLCL) with influence from mitochondrial membrane curvature. The protein contains domains and motifs involved in targeting, anchoring, and an active site for transacylase activity. Tafazzin activity affects many aspects of mitochondrial structure and function, including that of the electron transport chain, fission-fusion, as well as apoptotic signaling. TAZ mutations are implicated in Barth syndrome, an underdiagnosed and devastating disease that primarily affects male pediatric patients with a broad spectrum of disease pathologies that impact the cardiovascular, neuromuscular, metabolic, and hematologic systems.


Assuntos
Aciltransferases/genética , Síndrome de Barth/etiologia , Síndrome de Barth/genética , Síndrome de Barth/metabolismo , Cardiolipinas/genética , Mitocôndrias/genética , Fatores de Transcrição/genética , Animais , Apoptose/genética , Humanos , Transdução de Sinais/genética
9.
Am J Physiol Renal Physiol ; 318(1): F53-F66, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31657248

RESUMO

The mechanistic link between obesity and renal failure has been proposed to involve mitochondria reactive oxygen species generation and lipotoxicity. These pathological conditions make mitochondria of particular interest in the regulation of cell function and death by both apoptosis and autophagy. Therefore, this work was undertaken to investigate mitochondria function, autophagy, and apoptosis protein markers in the kidney from a rat model of intra-abdominal obesity and renal damage induced by a high-sucrose diet. Mitochondria from sucrose-fed (SF) kidneys in the presence of pyruvate-malate generated H2O2 at a higher rate than from control (79.81 ± 4.98 vs. 65.84 ± 1.95 pmol·min-1·mg protein-1). With succinate, the release of H2O2 was significantly higher compared with pyruvate-malate, and it remained higher in SF than in control mitochondria (146.4 ± 8.8 vs. 106.1 ± 5.9 pmol·min-1·mg protein-1). However, cytochrome c release from SF kidney mitochondria was lower than from control. In addition, cardiolipin, a mitochondria-specific phospholipid, was found increased in SF mitochondria due to the enhanced amount of both cardiolipin synthase and tafazzin. Cardiolipin was also found enriched with saturated and monounsaturated fatty acids, which are less susceptible to peroxidative stress involved in cytochrome c release. Furthermore, beclin-1 and light chain 3-B, as autophagy protein markers, and caspase-9, as apoptosis protein marker, were found decreased in SF kidneys. These results suggest that the decline of autophagy protein markers and the lack of apoptosis process could be a pathological mechanism of cell dysfunction leading to the progression of renal disease in SF rats.


Assuntos
Autofagia/fisiologia , Cardiolipinas/metabolismo , Sacarose na Dieta , Nefropatias/metabolismo , Rim/metabolismo , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Animais , Apoptose/fisiologia , Biomarcadores/metabolismo , Citocromos c/metabolismo , Peróxido de Hidrogênio/metabolismo , Masculino , Ratos , Ratos Wistar
10.
Artigo em Inglês | MEDLINE | ID: mdl-31884050

RESUMO

The mitochondrial phospholipid (CL) has been linked to mitochondrial and cellular functions. It has been postulated that the composition of CL is of impact for mitochondrial energy metabolism and cell proliferation. Although a correlation between CL composition and proliferation could be demonstrated for several cell types, evidence for a causal relationship remains obscure. Here, we applied two independent approaches, i) supplementation of fatty acids and ii) knock-out of the phospholipid remodeling enzyme tafazzin, to manipulate CL composition and analyzed the response on proliferation of C6 glioma cells. Both strategies caused substantial changes in the distribution of cellular fatty acids as well as in the distribution of fatty acids incorporated in CL that were accompanied by changes of the composition of molecular CL species. These changes did not correlate with cell proliferation. However, knock-out of tafazzin caused dramatic reduction in proliferation of C6 glioma cells independent of CL composition. The mechanism of tafazzin-dependent restriction of proliferation remains unclear. Among the various fatty acids administered only palmitic acid restricted cell proliferation by induction of cell death.


Assuntos
Aciltransferases/metabolismo , Neoplasias Encefálicas/metabolismo , Cardiolipinas/metabolismo , Glioma/metabolismo , Aciltransferases/genética , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ácidos Graxos/farmacologia , Fosfolipídeos/metabolismo , Ratos
11.
Oxid Med Cell Longev ; 2019: 9710208, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31827716

RESUMO

Peroxidation of cardiolipin (CL) in the inner mitochondrial membrane plays a key role in the development of various pathologies and, probably, aging. The four fatty acid tails of CL are usually polyunsaturated, which makes CL particularly sensitive to peroxidation. Peroxidation of CL is involved in the initiation of apoptosis, as well as in some other important cellular signaling chains. However, the studies of CL peroxidation are strongly limited by the lack of methods for its tracing in living cells. We have synthesized a new mitochondria-targeted fluorescent probe sensitive to lipid peroxidation (dubbed MitoCLox), where the BODIPY fluorophore, carrying a diene-containing moiety (as in the C11-BODIPY (581/591) probe), is conjugated with a triphenylphosphonium cation (TPP+) via a long flexible linker that contains two amide bonds. The oxidation of MitoCLox could be measured either as a decrease of absorbance at 588 nm or as an increase of fluorescence in the ratiometric mode at 520/590 nm (emission). In CL-containing liposomes, MitoCLox oxidation was induced by cytochrome c and developed in parallel with cardiolipin oxidation. TPP+-based mitochondria-targeted antioxidant SkQ1, in its reduced form, inhibited oxidation of MitoCLox concurrently with the peroxidation of cardiolipin. Molecular dynamic simulations of MitoCLox in a cardiolipin-containing membrane showed affinity of positively charged MitoCLox to negatively charged CL molecules; the oxidizable diene moiety of MitoCLox resided on the same depth as the cardiolipin lipid peroxides. We suggest that MitoCLox could be used for monitoring CL oxidation in vivo and, owing to its flexible linker, also serve as a platform for producing peroxidation sensors with affinity to particular lipids.


Assuntos
Cardiolipinas/química , Citocromos c/metabolismo , Corantes Fluorescentes/química , Peroxidação de Lipídeos , Mitocôndrias/metabolismo , Apoptose , Cardiolipinas/metabolismo , Fluorescência , Corantes Fluorescentes/metabolismo , Humanos , Lipossomos/metabolismo , Oxirredução , Espécies Reativas de Oxigênio/metabolismo
12.
Oxid Med Cell Longev ; 2019: 9186469, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31885824

RESUMO

Cardiolipin (CL) is a multifunctional dimeric phospholipid that physically interacts with electron transport chain complexes I, III, and IV, and ATP synthase (complex V). The enzyme ALCAT1 catalyzes the conversion of cardiolipin by incorporating polyunsaturated fatty acids into cardiolipin. The resulting CL species are said to be more susceptible to oxidative damage. This is thought to negatively affect the interaction of cardiolipin and electron transport chain complexes, leading to increased ROS production and mitochondrial dysfunction. Furthermore, it is discussed that ALCAT1 itself is upregulated due to oxidative stress. Here, we investigated the effects of overexpression of ALCAT1 under different metabolic conditions. ALCAT1 is located at the ER and mitochondria, probably at contact sites. We found that respiration stimulated by galactose supply promoted supercomplex assembly but also led to increased mitochondrial ROS levels. Endogeneous ALCAT1 protein expression levels showed a fairly high variability. Artificially induced ALCAT1 overexpression reduced supercomplex formation, further promoted ROS production, and prevented upregulation of coupled respiration. Taken together, our data suggest that the amount of the CL conversion enzyme ALCAT1 is critical for coupling mitochondrial respiration and metabolic plasticity.


Assuntos
1-Acilglicerol-3-Fosfato O-Aciltransferase/metabolismo , Cardiolipinas/metabolismo , Mitocôndrias/metabolismo , Complexos Multiproteicos/metabolismo , 1-Acilglicerol-3-Fosfato O-Aciltransferase/genética , Respiração Celular , Galactose/metabolismo , Células HeLa , Humanos , Potencial da Membrana Mitocondrial , Estresse Oxidativo , Multimerização Proteica/genética , Espécies Reativas de Oxigênio/metabolismo
13.
J Immunol Res ; 2019: 8380214, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31886311

RESUMO

Anticardiolipin antibody (ACA) is a kind of autoantibody and is one of the antiphospholipid antibodies (aPLs). Phospholipids with a negative charge on platelets and endothelial cell membranes are ACA target antigens. ACA is common in systemic lupus erythematosus and other autoimmune diseases and is closely associated with thrombosis, thrombocytopenia, and spontaneous abortion. In 1983, Harris established a method for detecting ACA, and research on the antibody has gained worldwide attention and has developed rapidly. For this review, we browsed articles that cover most of the ACA-related studies in the last 25 years and extracted influential ideas and conclusions in this field.


Assuntos
Anticorpos Anticardiolipina/imunologia , Autoantígenos/imunologia , Doenças Autoimunes/etiologia , Cardiolipinas/imunologia , Suscetibilidade a Doenças , Pesquisa , Doenças Autoimunes/diagnóstico , Doenças Autoimunes/metabolismo , Biomarcadores , Gerenciamento Clínico , Humanos
14.
Biochemistry (Mosc) ; 84(12): 1469-1483, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31870251

RESUMO

Age-related dysfunctions are accompanied by impairments in the mitochondrial morphology, activity of signaling pathway, and protein interactions. Cardiolipin is one of the most important phospholipids that maintains the curvature of the cristae and facilitates assembly and interaction of complexes and supercomplexes of the mitochondrial respiratory chain. The fatty acid composition of cardiolipin influences the biophysical properties of the membrane and, therefore, is crucial for the mitochondrial bioenergetics. The presence of unsaturated fatty acids in cardiolipin is the reason of its susceptibility to oxidative damage. Damaged cardiolipin undergoes remodeling by phospholipases, acyltransferases, and transacylases, creating a highly specific fatty acyl profile for each tissue. In this review, we discuss the variability of cardiolipin fatty acid composition in various species and different tissues of the same species, both in the norm and at various pathologies (e.g., age-related diseases, oxidative and traumatic stresses, knockouts/knockdowns of enzymes of the cardiolipin synthesis pathway). Progressive pathologies, including age-related ones, are accompanied by cardiolipin depletion and decrease in the efficiency of its remodeling, as well as the activation of an alternative way of pathological remodeling, which causes replacement of cardiolipin fatty acids with polyunsaturated ones (e.g., arachidonic or docosahexaenoic acids). Drugs or special diet can contribute to the partial restoration of the cardiolipin acyl profile to the one rich in fatty acids characteristic of an intact organ or tissue, thereby correcting the consequences of pathological or insufficient cardiolipin remodeling. In this regard, an urgent task of biomedicine is to study the mechanism of action of mitochondria-targeted antioxidants effective in the treatment of age-related pathologies and capable of accumulating not only in vitro, but also in vivo in the cardiolipin-enriched membrane fragments.


Assuntos
Envelhecimento/patologia , Cardiolipinas/metabolismo , Estresse Oxidativo , Animais , Antioxidantes/farmacologia , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo
15.
Oxid Med Cell Longev ; 2019: 3836186, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31885786

RESUMO

Chronic kidney disease (CKD) is highly incident and prevalent in the world. The death of patients with CKD is primarily due to cardiovascular disease. Renal transplantation (RT) emerges as the best management alternative for patients with CKD. However, the incidence of acute renal graft dysfunction is 11.8% of the related living donor and 17.4% of the cadaveric donor. Anticardiolipin antibodies (ACAs) or antiphospholipid antibodies (APAs) are important risk factors for acute renal graft dysfunction. The determination of ACA or APA to candidates for RT could serve as prognostic markers of early graft failure and would indicate which patients could benefit from anticoagulant therapy. Cardiolipin is a fundamental molecule that plays an important role in the adequate conformation of the mitochondrial cristae and the correct assembly of the mitochondrial respiratory supercomplexes and other proteins essential for proper mitochondrial function. Cardiolipin undergoes a nonrandom oxidation process by having pronounced specificity unrelated to the polyunsaturation pattern of its acyl groups. Accumulation of hydroxyl derivatives and cardiolipin hydroperoxides has been observed in the affected tissues, and recent studies showed that oxidation of cardiolipin is carried out by a cardiolipin-specific peroxidase activity of cardiolipin-bound cytochrome c. Cardiolipin could be responsible for the proapoptotic production of death signals. Cardiolipin modulates the production of energy and participates in inflammation, mitophagy, and cellular apoptosis. The determination of cardiolipin or its antibodies is an attractive therapeutic, diagnostic target in RT and kidney diseases.


Assuntos
Cardiolipinas/imunologia , Transplante de Rim , Mitocôndrias/metabolismo , Anticorpos/sangue , Cardiolipinas/metabolismo , Rejeição de Enxerto/etiologia , Humanos , Mitofagia , Estresse Oxidativo , Insuficiência Renal Crônica/terapia , Fatores de Risco
16.
Anal Bioanal Chem ; 411(30): 8123-8131, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31754767

RESUMO

Collision cross section (CCS) values are descriptors of the 3D structure of ions which can be determined by ion mobility spectrometry (IMS). Currently, most lipidomic studies involving CCS value determination concern eukaryote samples (e.g. human, bovine) and to a lower extent prokaryote samples (e.g. bacteria). Here, we report CCS values obtained from traveling wave ion mobility spectrometry (TWCCSN2) measurements from the bacterial membrane of Pseudomonas aeruginosa-a bacterium ranked as priority 1 for the R&D of new antibiotics by the World Health Organization. In order to cover the lack of reference compounds which could cover the m/z and CCS ranges of the membrane lipids of P. aeruginosa, three calibrants (polyalanine, dextran and phospholipids) were used for the TWCCSN2 calibration. A shift from the published lipid CCS values was systematically observed (ΔCCS% up to 9%); thus, we proposed a CCS correction strategy. This correction strategy allowed a reduction in the shift (ΔCCS%) between our measurements and published values to less than 2%. This correction was then applied to determine the CCS values of Pseudomonas aeruginosa lipids which have not been published yet. As a result, 32 TWCCSN2 values for [M+H]+ ions and 24 TWCCSN2 values for [M-H]- ions were obtained for four classes of phospholipids (phosphatidylethanolamines (PE), phosphatidylcholines (PC), phosphatidylglycerols (PG) and diphosphatidylglycerols-known as cardiolipins (CL)). Graphical abstract.


Assuntos
Cardiolipinas/análise , Espectrometria de Mobilidade Iônica/métodos , Espectrometria de Massas/métodos , Fosfolipídeos/análise , Calibragem
17.
Oxid Med Cell Longev ; 2019: 2901057, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31781330

RESUMO

Cardiolipin interacts with many proteins of the mitochondrial inner membrane and, together with cytochrome C and creatine kinase, activates them. It can be considered as an integrating factor for components of the mitochondrial respiratory chain, which provides for an efficient transfer of electrons and protons. The major, if not the only, factor of cardiolipin maturation is tafazzin. Variations of isoform proportions of this enzyme can cause severe diseases such as Barth syndrome. Using bioinformatic methods, we have found conserved C-terminal regions in many tafazzin isoforms and identified new mammalian species that acquired exon 5 as well as rare occasions of intron retention between exons 8 and 9. The regions in the C-terminal part arise from frameshifts relative to the full-length TAZ transcript after skipping exon 9 or retention of the intron between exons 10 and 11. These modifications demonstrate specific distribution among the orders of mammals. The dependence of the species maximum lifespan, body weight, and mitochondrial metabolic rate on the modifications has been demonstrated. Arguably, unconventional tafazzin isoforms provide for the optimal balance between the increased biochemical activity of mitochondria (resulting from specific environmental or nutritional conditions) and lifespan maintenance; and the functional role of such isoforms is linked to the modification of the primary and secondary structures at their C-termini.


Assuntos
Síndrome de Barth/metabolismo , Cardiolipinas/metabolismo , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Fatores de Transcrição/metabolismo , Animais , Síndrome de Barth/genética , Síndrome de Barth/patologia , Cardiolipinas/genética , Transporte de Elétrons/genética , Humanos , Mitocôndrias/genética , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Fatores de Transcrição/genética
18.
Rinsho Shinkeigaku ; 59(10): 662-665, 2019 Oct 26.
Artigo em Japonês | MEDLINE | ID: mdl-31564704

RESUMO

A 55-year-old man presented with recurrent brain infarction which had increased multifocally mainly in the cerebral white matter over the course of one year. Antibodies associated with antiphospholipid syndrome (APS) were initially negative. The patient was admitted to our department because of the thickened meninges shown on gadolinium enhanced brain MRI, mimicking hypertrophic pachymeningitis. However, blood and cerebrospinal fluid analysis revealed no significant inflammatory changes. On histopathological examination of the biopsied meninges, the arachnoid membrane was thickened with fibrosis, and arachnoidal microvessels were enlarged without significant inflammatory changes. The dura mater was not thickened, and no inflammation or microvessel enlargement were revealed. Finally, serum IgG anticardiolipin antibody testing was positive twice at an interval of more than 12 weeks, confirming the diagnosis of APS. Since initiating antithrombotic therapy with warfarin, brain infarction has not recurred. Without inflammation in the arachnoid membrane, the congestion of blood flow caused by thrombosis of microvessels in the arachnoid membrane might have increased the thickness of the arachnoid membrane.


Assuntos
Síndrome Antifosfolipídica/complicações , Síndrome Antifosfolipídica/diagnóstico , Aracnoide-Máter/patologia , Infarto Cerebral/etiologia , Anticoagulantes/administração & dosagem , Síndrome Antifosfolipídica/tratamento farmacológico , Aracnoide-Máter/irrigação sanguínea , Aracnoide-Máter/diagnóstico por imagem , Biomarcadores/sangue , Cardiolipinas/imunologia , Infarto Cerebral/prevenção & controle , Humanos , Hipertrofia/etiologia , Imunoglobulina G/sangue , Imagem por Ressonância Magnética , Masculino , Microvasos , Pessoa de Meia-Idade , Recidiva , Trombose/complicações , Resultado do Tratamento , Varfarina/administração & dosagem
19.
J Exp Clin Cancer Res ; 38(1): 436, 2019 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-31665044

RESUMO

BACKGROUND: All-trans-retinoic-acid (ATRA) is a promising agent in the prevention/treatment of breast-cancer. There is growing evidence that reprogramming of cellular lipid metabolism contributes to malignant transformation and progression. Lipid metabolism is implicated in cell differentiation and metastatic colonization and it is involved in the mechanisms of sensitivity/resistance to different anti-tumor agents. The role played by lipids in the anti-tumor activity of ATRA has never been studied. METHODS: We used 16 breast cancer cell-lines whose degree of sensitivity to the anti-proliferative action of ATRA is known. We implemented a non-oriented mass-spectrometry based approach to define the lipidomic profiles of each cell-line grown under basal conditions and following treatment with ATRA. To complement the lipidomic data, untreated and retinoid treated cell-lines were also subjected to RNA-sequencing to define the perturbations afforded by ATRA on the whole-genome gene-expression profiles. The number and functional activity of mitochondria were determined in selected ATRA-sensitive and -resistant cell-lines. Bio-computing approaches were used to analyse the high-throughput lipidomic and transcriptomic data. RESULTS: ATRA perturbs the homeostasis of numerous lipids and the most relevant effects are observed on cardiolipins, which are located in the mitochondrial inner membranes and play a role in oxidative-phosphorylation. ATRA reduces the amounts of cardiolipins and the effect is associated with the growth-inhibitory activity of the retinoid. Down-regulation of cardiolipins is due to a reduction of mitochondria, which is caused by an ATRA-dependent decrease in the expression of nuclear genes encoding mitochondrial proteins. This demonstrates that ATRA anti-tumor activity is due to a decrease in the amounts of mitochondria causing deficits in the respiration/energy-balance of breast-cancer cells. CONCLUSIONS: The observation that ATRA anti-proliferative activity is caused by a reduction in the respiration and energy balance of the tumor cells has important ramifications for the therapeutic action of ATRA in breast cancer. The study may open the way to the development of rational therapeutic combinations based on the use of ATRA and anti-tumor agents targeting the mitochondria.


Assuntos
Neoplasias da Mama/metabolismo , Cardiolipinas/metabolismo , Perfilação da Expressão Gênica/métodos , Mitocôndrias/metabolismo , Tretinoína/farmacologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Lipidômica/métodos , Espectrometria de Massas , Mitocôndrias/efeitos dos fármacos , Fosforilação Oxidativa/efeitos dos fármacos , Análise de Célula Única , Sequenciamento Completo do Exoma
20.
Exp Mol Pathol ; 111: 104318, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31614130

RESUMO

Ketamine is widely used both as anesthetic and abuse drug. In this study, we investigated the effects of a wide range of ketamine concentrations (100-500-1000 µM) on calcium mobilization and the induction of cell death in undifferentiated PC12 cells, 24 h after treatment. Calcium mobilization was measured as the percentage of fluorescence one minute after depolarization by flow cytometry. For the kinetic changes in [Ca2+]c, fluorescence microscopy with Live Imaging was used with a resolution time of 0.87 s (exposure time: 20 ms). Fluo-4 AM was used for both methods. Flow cytometry using TMRE, NAO, and Annexin V-FITC/PI probes were employed for the evaluation of mitochondrial membrane potential (ΔΨm), cardiolipin content and type of cell death respectively. Fluorescence microscopy was used for the evaluation of DNA fragmentation by TUNEL assay with dUTP-conjugated FITC. Results obtained by flow cytometry showed a clear increment in cell response to depolarization after addition of 50 mM and 70 mM KCl in PC12 cells. Simultaneously, cells treated with 100 µM and 500 µM ketamine during 24 h, induced a decreased response to depolarization as compared with control cells. In addition, 1000 µM ketamine induced a similar increase in Fluo4AM fluorescence either after addition of 50 or 70 mM KCl. The kinetic assays showed that after 100 mM KCl, cells pre-treated with ketamine showed a marked decrease in [Ca2+]c as compared with control cells. In the case of 1000 µM ketamine treatment, an increased and sustained [Ca2+]c was observed along the whole assay, indicating a cell disability to maintain calcium homeostasis. Associated with these cytosolic calcium alterations, mitochondrial depolarization, cardiolipin depletion and alteration in Bax protein expression were observed after ketamine treatment. Our data demonstrate that ketamine action in these cells seems to be independent from NMDAR, as observed by the absence of glutamate­calcium response. Acute disturbance in [Ca2+]c could be mediated by the inhibition of VDCCs as part of the molecular mechanism of ketamine cytotoxicity leading to mitochondrial dysfunction and cell death by apoptosis and necrosis.


Assuntos
Canais de Cálcio/metabolismo , Ketamina/farmacologia , Analgésicos/administração & dosagem , Analgésicos/farmacologia , Animais , Sinalização do Cálcio/efeitos dos fármacos , Cardiolipinas/metabolismo , Morte Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ketamina/administração & dosagem , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Células PC12 , Cloreto de Potássio/farmacologia , Ratos , Proteína X Associada a bcl-2/metabolismo
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