RESUMO
Coenzyme Q10 (CoQ10) is an important cofactor and antioxidant for numerous cellular processes, and its deficiency has been linked to human disorders including mitochondrial disease, heart failure, Parkinson's disease, and hypertension. Unfortunately, treatment with exogenous CoQ10 is often ineffective, likely due to its extreme hydrophobicity and high molecular weight. Here, we show that less hydrophobic CoQ species with shorter isoprenoid tails can serve as viable substitutes for CoQ10 in human cells. We demonstrate that CoQ4 can perform multiple functions of CoQ10 in CoQ-deficient cells at markedly lower treatment concentrations, motivating further investigation of CoQ4 as a supplement for CoQ10 deficiencies. In addition, we describe the synthesis and evaluation of an initial set of compounds designed to target CoQ4 selectively to mitochondria using triphenylphosphonium. Our results indicate that select versions of these compounds can successfully be delivered to mitochondria in a cell model and be cleaved to produce CoQ4, laying the groundwork for further development.
Assuntos
Ataxia , Mitocôndrias , Doenças Mitocondriais , Debilidade Muscular , Ubiquinona , Humanos , Mitocôndrias/enzimologia , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Debilidade Muscular/enzimologia , Debilidade Muscular/genética , Ubiquinona/análogos & derivados , Ubiquinona/deficiência , Células Hep G2RESUMO
Mitochondrial fatty acid oxidation disorders (FAODs) are caused by defects in ß-oxidation enzymes, including very long-chain acyl-CoA dehydrogenase (VLCAD), trifunctional protein (TFP), carnitine palmitoyltransferase-2 (CPT2), carnitine-acylcarnitine translocase (CACT) and others. During prolonged fasting, infection, or exercise, patients with FAODs present with hypoglycemia, rhabdomyolysis, cardiomyopathy, liver dysfunction, and occasionally sudden death. This article describes the diagnosis, newborn screening, and treatment of long-chain FAODs with a focus on VLCAD deficiency. VLCAD deficiency is generally classified into three phenotypes based on onset time, but the classification should be comprehensively determined based on genotype, residual enzyme activity, and clinical course, due to a lack of apparent genotype-phenotype correlation. With the expansion of newborn screening for FAODs, several issues have arisen, such as missed detection, overdiagnosis (including detection of benign/asymptomatic type), and poor prognosis of the neonatal-onset form. Meanwhile, dietary management and restriction of exercise have been unnecessary for patients with the benign/asymptomatic type of VLCAD deficiency with a high fatty acid oxidation flux score. Although L-carnitine therapy for VLCAD/TFP deficiency has been controversial, supplementation with L-carnitine may be accepted for CPT2/CACT and multiple acyl-CoA dehydrogenase deficiencies. Recently, a double-blind, randomized controlled trial of triheptanoin (seven-carbon fatty acid triglyceride) versus trioctanoin (regular medium-chain triglyceride) was conducted and demonstrated improvement of cardiac functions on triheptanoin. Additionally, although the clinical efficacy of bezafibrate remains controversial, a recent open-label clinical trial showed efficacy of this drug in improving quality of life. These drugs may be promising for the treatment of FAODs, though further studies are required.
Assuntos
Acil-CoA Desidrogenase de Cadeia Longa/deficiência , Ácidos Graxos/metabolismo , Hipolipemiantes/uso terapêutico , Erros Inatos do Metabolismo Lipídico/diagnóstico , Erros Inatos do Metabolismo Lipídico/tratamento farmacológico , Programas de Rastreamento , Doenças Mitocondriais/diagnóstico , Doenças Mitocondriais/tratamento farmacológico , Doenças Musculares/diagnóstico , Doenças Musculares/tratamento farmacológico , Síndrome Congênita de Insuficiência da Medula Óssea , Gerenciamento Clínico , Humanos , Erros Inatos do Metabolismo Lipídico/enzimologia , Doenças Mitocondriais/enzimologia , Doenças Musculares/enzimologiaRESUMO
Mitochondrial complex I (CI) deficiency is the most frequent cause of oxidative phosphorylation (OXPHOS) disorders in humans. In order to benchmark the effects of CI deficiency on mitochondrial bioenergetics and dynamics, respiratory chain (RC) and endoplasmic reticulum (ER)-mitochondria communication, and superoxide production, fibroblasts from patients with mutations in the ND6, NDUFV1 or ACAD9 genes were analyzed. Fatty acid metabolism, basal and maximal respiration, mitochondrial membrane potential, and ATP levels were decreased. Changes in proteins involved in mitochondrial dynamics were detected in various combinations in each cell line, while variable changes in RC components were observed. ACAD9 deficient cells exhibited an increase in RC complex subunits and DDIT3, an ER stress marker. The level of proteins involved in ER-mitochondria communication was decreased in ND6 and ACAD9 deficient cells. |ΔΨ| and cell viability were further decreased in all cell lines. These findings suggest that disruption of mitochondrial bioenergetics and dynamics, ER-mitochondria crosstalk, and increased superoxide contribute to the pathophysiology in patients with ACAD9 deficiency. Furthermore, treatment of ACAD9 deficient cells with JP4-039, a novel mitochondria-targeted reactive oxygen species, electron and radical scavenger, decreased superoxide level and increased basal and maximal respiratory rate, identifying a potential therapeutic intervention opportunity in CI deficiency.
Assuntos
Acil-CoA Desidrogenases/genética , Complexo I de Transporte de Elétrons/deficiência , Fibroblastos/enzimologia , Doenças Mitocondriais/genética , NADH Desidrogenase/genética , Espécies Reativas de Oxigênio/metabolismo , Acil-CoA Desidrogenases/deficiência , Trifosfato de Adenosina/agonistas , Trifosfato de Adenosina/biossíntese , Transporte de Elétrons/efeitos dos fármacos , Transporte de Elétrons/genética , Complexo I de Transporte de Elétrons/genética , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Retículo Endoplasmático/patologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Sequestradores de Radicais Livres/farmacologia , Expressão Gênica , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/enzimologia , Mitocôndrias/patologia , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/patologia , NADH Desidrogenase/deficiência , Óxidos de Nitrogênio/farmacologia , Fosforilação Oxidativa/efeitos dos fármacos , Cultura Primária de Células , Espécies Reativas de Oxigênio/antagonistas & inibidoresRESUMO
Methylmalonic aciduria (MMA) is a disorder of organic acid metabolism resulting from a functional defect of the mitochondrial enzyme, methylmalonyl-CoA mutase (MCM). The main treatments for MMA patients are dietary restriction of propiogenic amino acids and carnitine supplementation. Liver or combined liver/kidney transplantation has been used to treat those with the most severe clinical manifestations. Thus, therapies are necessary to help improve quality of life and prevent liver, renal and neurological complications. Previously, we successfully used the TAT-MTS-Protein approach for replacing a number of mitochondrial-mutated proteins. In this targeted system, TAT, an 11 a.a peptide, which rapidly and efficiently can cross biological membranes, is fused to a mitochondrial targeting sequence (MTS), followed by the mitochondrial mature protein which sends the protein into the mitochondria. In the mitochondria, the TAT-MTS is cleaved off and the native protein integrates into its natural complexes and is fully functional. In this study, we used heterologous MTSs of human, nuclear-encoded mitochondrial proteins, to target the human MCM protein into the mitochondria. All fusion proteins reached the mitochondria and successfully underwent processing. Treatment of MMA patient fibroblasts with these fusion proteins restored mitochondrial activity such as ATP production, mitochondrial membrane potential and oxygen consumption, indicating the importance of mitochondrial function in this disease. Treatment with the fusion proteins enhanced cell viability and most importantly reduced MMA levels. Treatment also enhanced albumin and urea secretion in a CRISPR/Cas9-engineered HepG2 MUT (-/-) liver cell line. Therefore, we suggest using this TAT-MTS-Protein approach for the treatment of MMA.
Assuntos
Trifosfato de Adenosina/biossíntese , Fibroblastos/enzimologia , Produtos do Gene tat/genética , Metilmalonil-CoA Mutase/genética , Mitocôndrias/enzimologia , Proteínas Recombinantes de Fusão/genética , Erros Inatos do Metabolismo dos Aminoácidos/enzimologia , Erros Inatos do Metabolismo dos Aminoácidos/genética , Erros Inatos do Metabolismo dos Aminoácidos/patologia , Erros Inatos do Metabolismo dos Aminoácidos/terapia , Sistemas CRISPR-Cas , Escherichia coli/genética , Escherichia coli/metabolismo , Fibroblastos/patologia , Expressão Gênica , Produtos do Gene tat/metabolismo , Terapia Genética/métodos , Células Hep G2 , Humanos , Fígado/enzimologia , Fígado/patologia , Potencial da Membrana Mitocondrial , Ácido Metilmalônico/metabolismo , Metilmalonil-CoA Mutase/metabolismo , Mitocôndrias/patologia , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Doenças Mitocondriais/patologia , Doenças Mitocondriais/terapia , Plasmídeos/química , Plasmídeos/metabolismo , Cultura Primária de Células , Engenharia de Proteínas/métodos , Sinais Direcionadores de Proteínas/genética , Proteínas Recombinantes de Fusão/metabolismo , TransfecçãoRESUMO
In recent years, the analytical determination of coenzyme Q10 (CoQ10) has gained importance in clinical diagnosis and in pharmaceutical quality control. CoQ10 is an important cofactor in the mitochondrial respiratory chain and a potent endogenous antioxidant. CoQ10 deficiency is often associated with numerous diseases and patients with these conditions may benefit from administration of supplements of CoQ10. In this regard, it has been observed that the best benefits are obtained when CoQ10 deficiency is diagnosed and treated early. Therefore, it is of great value to develop analytical methods for the detection and quantification of CoQ10 in this type of disease. The methods above mentioned should be simple enough to be used in routine clinical laboratories as well as in quality control of pharmaceutical formulations containing CoQ10. Here, we discuss the advantages and disadvantages of different methods of CoQ10 analysis.
Assuntos
Ubiquinona/análogos & derivados , Ataxia/diagnóstico , Ataxia/enzimologia , Cromatografia Líquida de Alta Pressão , Eletroforese Capilar , Humanos , Doenças Mitocondriais/diagnóstico , Doenças Mitocondriais/enzimologia , Debilidade Muscular/diagnóstico , Debilidade Muscular/enzimologia , Preparações Farmacêuticas/química , Espectrofotometria , Ubiquinona/análise , Ubiquinona/sangue , Ubiquinona/química , Ubiquinona/deficiência , Ubiquinona/isolamento & purificaçãoRESUMO
Accumulating research has shown that chronic D-galactose (D-gal) exposure induces symptoms similar to natural aging in animals. Therefore, rodents chronically exposed to D-gal are increasingly used as a model for aging and delay-of-aging pharmacological research. Mitochondrial dysfunction is thought to play a vital role in aging and age-related diseases; however, whether mitochondrial dysfunction plays a significant role in mice exposed to D-gal remains unknown. In the present study, we investigated cognitive dysfunction, locomotor activity, and mitochondrial dysfunction involved in D-gal exposure in mice. We found that D-gal exposure (125 mg/kg/day, 8 weeks) resulted in a serious impairment in grip strength in mice, whereas spatial memory and locomotor coordination remained intact. Interestingly, muscular mitochondrial complex I deficiency occurred in the skeletal muscle of mice exposed to D-gal. Mitochondrial ultrastructure abnormality was implicated as a contributing factor in D-gal-induced muscular impairment. Moreover, three combinations (A, B, and C) of nutrients applied in this study effectively reversed D-gal-induced muscular impairment. Nutrient formulas B and C were especially effective in reversing complex I dysfunction in both skeletal muscle and heart muscle. These findings suggest the following: (1) chronic exposure to D-gal first results in specific muscular impairment in mice, rather than causing general, premature aging; (2) poor skeletal muscle strength induced by D-gal might be due to the mitochondrial dysfunction caused by complex I deficiency; and (3) the nutrient complexes applied in the study attenuated the skeletal muscle impairment, most likely by improving mitochondrial function.
Assuntos
Complexo I de Transporte de Elétrons/deficiência , Galactose/efeitos adversos , Doenças Mitocondriais/etiologia , Músculo Esquelético/metabolismo , Animais , Modelos Animais de Doenças , Complexo I de Transporte de Elétrons/metabolismo , Feminino , Galactose/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/enzimologia , Mitocôndrias/metabolismo , Doenças Mitocondriais/dietoterapia , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/metabolismo , Músculo Esquelético/enzimologiaRESUMO
Human COQ6 encodes a monooxygenase which is responsible for the C5-hydroxylation of the quinone ring of coenzyme Q (CoQ). Mutations in COQ6 cause primary CoQ deficiency, a condition responsive to oral CoQ10 supplementation. Treatment is however still problematic given the poor bioavailability of CoQ10. We employed S. cerevisiae lacking the orthologous gene to characterize the two different human COQ6 isoforms and the mutations found in patients. COQ6 isoform a can partially complement the defective yeast, while isoform b, which lacks part of the FAD-binding domain, is inactive but partially stable, and could have a regulatory/inhibitory function in CoQ10 biosynthesis. Most mutations identified in patients, including the frameshift Q461fs478X mutation, retain residual enzymatic activity, and all patients carry at least one hypomorphic allele, confirming that the complete block of CoQ biosynthesis is lethal. These mutants are also partially stable and allow the assembly of the CoQ biosynthetic complex. In fact treatment with two hydroxylated analogues of 4-hydroxybenzoic acid, namely, vanillic acid or 3-4-hydroxybenzoic acid, restored the respiratory growth of yeast Δcoq6 cells expressing the mutant huCOQ6-isoa proteins. These compounds, and particularly vanillic acid, could therefore represent an interesting therapeutic option for COQ6 patients.
Assuntos
Aminobenzoatos/farmacologia , Hidroxibenzoatos/farmacologia , Mutação , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/efeitos dos fármacos , Ubiquinona/genética , Ácido Vanílico/farmacologia , Sequência de Aminoácidos , Ataxia/tratamento farmacológico , Ataxia/enzimologia , Ataxia/genética , Expressão Gênica , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/enzimologia , Mitocôndrias/genética , Doenças Mitocondriais/tratamento farmacológico , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Modelos Moleculares , Dados de Sequência Molecular , Debilidade Muscular/tratamento farmacológico , Debilidade Muscular/enzimologia , Debilidade Muscular/genética , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Ubiquinona/análogos & derivados , Ubiquinona/química , Ubiquinona/deficiência , Ubiquinona/metabolismoRESUMO
Nutrition has been highlighted as a potential factor in Alzheimer's disease (AD) risk and decline and has been investigated as a therapeutic target. Broad-based combination diet therapies have the potential to simultaneously effect numerous protective and corrective processes, both directly (e.g., neuroprotection) and indirectly (e.g., improved vascular health). Here we administered either normal mouse chow with a broad-based nutritional supplement or mouse chow alone to aged male and female 3xTg mice and wildtype (WT) controls. After approximately 4 months of feeding, mice were given a battery of cognitive tasks and then injected with a radiolabeled glucose analog. Brains were assessed for differences in regional glucose uptake and mitochondrial cytochrome oxidase activity, AD pathology, and inflammatory markers. Supplementation induced behavioral changes in the 3xTg, but not WT, mice, and the mode of these changes was influenced by sex. Subsequent analyses indicated that differential response to supplementation by male and female 3xTg mice highlighted brain regional strategies for the preservation of function. Several regions involved have been shown to mediate responses to steroid hormones, indicating a mechanism for sex-based vulnerability. Thus, these findings may have broad implications for the human response to future therapeutics.
Assuntos
Doença de Alzheimer/dietoterapia , Doença de Alzheimer/psicologia , Suplementos Nutricionais , Doenças Mitocondriais/dietoterapia , Fenômenos Fisiológicos da Nutrição , Doença de Alzheimer/diagnóstico por imagem , Amiloidose/patologia , Animais , Cognição/fisiologia , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Fluordesoxiglucose F18 , Proteína Glial Fibrilar Ácida/metabolismo , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Inflamação/patologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Doenças Mitocondriais/enzimologia , Extratos Vegetais/uso terapêutico , Desempenho Psicomotor/fisiologia , Cintilografia , Compostos Radiofarmacêuticos , Caracteres Sexuais , Sinaptofisina/metabolismo , Proteínas tau/metabolismoRESUMO
We describe a 22-year-old male who developed severe hypoglycemia and lethargy during an acute illness at 4 months of age and subsequently grew and developed normally. At age 4 years he developed recurrent vomiting with mild hyperammonemia and dehydration requiring frequent hospitalizations. Glutaric aciduria Type II was suspected based upon biochemical findings and managed with cornstarch, carnitine and riboflavin supplements. He did not experience metabolic crises between ages 4-12 years. He experienced recurrent vomiting, mild hyperammonemia, and generalized weakness associated with acute illnesses and growth spurts. At age 18 years, he developed exercise intolerance and proximal muscle weakness leading to the identification of multiple acyl-CoA dehydrogenase and complex II/III deficiencies in both skeletal muscle and liver. Subsequent molecular characterization of the ETFDH gene revealed novel heterozygous mutations, p.G274X:c.820 G > T (exon 7) and p.P534L: c.1601 C > T (exon 12), the latter within the iron sulfur-cluster and predicted to affect ubiquinone reductase activity of ETFDH and the docking of ETF to ETFDH. Our case supports the concept of a structural interaction between ETFDH and other enzyme partners, and suggests that the conformational change upon ETF binding to ETFDH may play a key role in linking ETFDH to II/III super-complex formation.
Assuntos
Complexo III da Cadeia de Transporte de Elétrons/deficiência , Complexo II de Transporte de Elétrons/deficiência , Flavoproteínas Transferidoras de Elétrons/genética , Proteínas Ferro-Enxofre/genética , Fígado/enzimologia , Erros Inatos do Metabolismo/genética , Doenças Mitocondriais/genética , Deficiência Múltipla de Acil Coenzima A Desidrogenase/genética , Músculo Esquelético/enzimologia , Mutação , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Biomarcadores/sangue , Biomarcadores/urina , Análise Mutacional de DNA , Complexo II de Transporte de Elétrons/química , Complexo II de Transporte de Elétrons/genética , Complexo II de Transporte de Elétrons/metabolismo , Complexo III da Cadeia de Transporte de Elétrons/química , Complexo III da Cadeia de Transporte de Elétrons/genética , Complexo III da Cadeia de Transporte de Elétrons/metabolismo , Flavoproteínas Transferidoras de Elétrons/química , Flavoproteínas Transferidoras de Elétrons/deficiência , Éxons , Predisposição Genética para Doença , Heterozigoto , Humanos , Proteínas Ferro-Enxofre/química , Proteínas Ferro-Enxofre/deficiência , Masculino , Erros Inatos do Metabolismo/diagnóstico , Erros Inatos do Metabolismo/enzimologia , Doenças Mitocondriais/diagnóstico , Doenças Mitocondriais/enzimologia , Simulação de Acoplamento Molecular , Deficiência Múltipla de Acil Coenzima A Desidrogenase/diagnóstico , Deficiência Múltipla de Acil Coenzima A Desidrogenase/enzimologia , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/química , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/deficiência , Fenótipo , Ligação Proteica , Conformação Proteica , Adulto JovemRESUMO
Treatment recommendations in mitochondrial fatty acid oxidation (FAO) defects are diverse. With implementation of newborn screening and identification of asymptomatic patients, it is necessary to define whom to treat and how strictly. We here discuss critical questions that are currently under debate. For some asymptomatic long-chain defects, long-chain fat restriction plays a minor role, and a normal diet may be introduced. For patients presenting only with myopathic symptoms, e.g., during exercise, treatment may be adapted to energy demand. As a consequence, patients with exercise-induced myopathy may be able to return to normal activity when provided with medium-chain triglycerides (MCT) prior to exercise. There is no need to limit participation in sports. Progression of retinopathy in disorders of the mitochondrial trifunctional protein complex is closely associated with hydroxyacylcarnitine accumulation. A strict low-fat diet with MCT supplementation is recommended to slow or prevent progression of chorioretinopathy. Additional docosahexanoic acid does not prevent the decline in retinal function but does promote nonspecific improvement in visual acuity and is recommended. There is no evidence that L-carnitine supplementation is beneficial. Thus, supplementation with L-carnitine in a newborn identified by screening with either a medium-chain or long-chain defect is not supported. With respect to the use of the odd-chain medium-chain triglyceride triheptanoin in myopathic phenotypes, randomized trials are needed to establish whether triheptanoin is more effective than even-chain MCT. With increasing pathophysiological knowledge, new treatment options have been identified and are being clinically evaluated. These include the use of bezafibrates in myopathic long-chain defects.
Assuntos
Metabolismo Energético , Ácidos Graxos/metabolismo , Erros Inatos do Metabolismo Lipídico/terapia , Mitocôndrias/enzimologia , Doenças Mitocondriais/terapia , Metabolismo Energético/genética , Medicina Baseada em Evidências , Genótipo , Humanos , Erros Inatos do Metabolismo Lipídico/complicações , Erros Inatos do Metabolismo Lipídico/diagnóstico , Erros Inatos do Metabolismo Lipídico/enzimologia , Erros Inatos do Metabolismo Lipídico/genética , Doenças Mitocondriais/complicações , Doenças Mitocondriais/diagnóstico , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Oxirredução , Fenótipo , Guias de Prática Clínica como Assunto , Resultado do TratamentoRESUMO
Mouse models have been designed for a number of fatty acid oxidation defects. Studies in these mouse models have demonstrated that different pathogenetic mechanisms play a role in the pathophysiology of defects of fatty acid oxidation. Supplementation with L-carnitine does not prevent low tissue carnitine levels and induces acylcarnitine production having potentially toxic effects, as presented in very-long-chain acyl-CoA dehydrogenase (VLCAD)-deficient mice. Energy deficiency appears to be an important mechanism in the development of cardiomyopathy and skeletal myopathy in fatty acid oxidation defects and is also the underlying mechanism of cold intolerance. Hypoglycemia as one major clinical sign in all fatty acid oxidation defects occurs due to a reduced hepatic glucose output and an enhanced peripheral glucose uptake rather than to transcriptional changes that are also observed simultaneously, as presented in medium-chain acyl-CoA dehydrogenase (MCAD)-deficient mice. There are reports that an impaired fatty acid oxidation also plays a role in intrauterine life. The embryonic loss demonstrated for some enzyme defects in the mouse supports this hypothesis. However, the exact mechanisms are unknown. This observation correlates to maternal hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome, as observed in pregnancies carrying a long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD)-deficient fetus. Synergistic heterozygosity has been shown in isolated patients and in mouse models to be associated with clinical phenotypes common to fatty acid oxidation disorders. Synergistic mutations may also modulate severity of the clinical phenotype and explain in part clinical heterogeneity of fatty acid oxidation defects. In summary, knowledge about the different pathogenetic mechanisms and the resulting pathophysiology allows the development of specific new therapies.
Assuntos
Metabolismo Energético , Ácidos Graxos/metabolismo , Erros Inatos do Metabolismo Lipídico/fisiopatologia , Mitocôndrias/enzimologia , Doenças Mitocondriais/fisiopatologia , Animais , Modelos Animais de Doenças , Metabolismo Energético/genética , Genótipo , Humanos , Erros Inatos do Metabolismo Lipídico/complicações , Erros Inatos do Metabolismo Lipídico/enzimologia , Erros Inatos do Metabolismo Lipídico/genética , Camundongos , Doenças Mitocondriais/complicações , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Oxirredução , Fenótipo , Especificidade da EspécieRESUMO
Three of our research efforts are reviewed, which suggest that optimizing metabolism will delay aging and the diseases of aging in humans. (1) Research on delay of the mitochondrial decay of aging by supplementing rats with lipoic acid and acetyl carnitine. (2) The triage theory, which posits that modest micronutrient deficiencies (common in much of the population) accelerate molecular aging, including mitochondrial decay, and supportive evidence, including an analysis in depth of vitamin K, that suggests the importance of achieving optimal micronutrient intake for longevity. (3) The finding that decreased enzyme binding constants (increased Km) for coenzymes (or substrates) can result from protein deformation and loss of function due to loss of membrane fluidity with age, or to polymorphisms or mutation. The loss of enzyme function can be ameliorated by high doses of a B vitamin, which raises coenzyme levels, and indicates the importance of understanding the effects of age, or polymorphisms, on micronutrient requirements.
Assuntos
Envelhecimento/metabolismo , Senescência Celular/efeitos dos fármacos , Suplementos Nutricionais , Micronutrientes/administração & dosagem , Mitocôndrias/efeitos dos fármacos , Doenças Mitocondriais/prevenção & controle , Acetilcarnitina/administração & dosagem , Fatores Etários , Envelhecimento/patologia , Animais , Coenzimas/administração & dosagem , Enzimas/metabolismo , Humanos , Longevidade , Micronutrientes/deficiência , Micronutrientes/metabolismo , Mitocôndrias/enzimologia , Mitocôndrias/patologia , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/patologia , Ácido Tióctico/administração & dosagem , Vitamina K/administração & dosagemAssuntos
Predisposição Genética para Doença/genética , Transtornos do Metabolismo dos Lipídeos/enzimologia , Transtornos do Metabolismo dos Lipídeos/genética , Músculo Esquelético/enzimologia , Doenças Musculares/enzimologia , Doenças Musculares/genética , Suplementos Nutricionais , Terapia por Exercício , Humanos , Transtornos do Metabolismo dos Lipídeos/terapia , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Doenças Mitocondriais/fisiopatologia , Músculo Esquelético/patologia , Músculo Esquelético/fisiopatologia , Doenças Musculares/terapia , Países Baixos , Farmacologia/tendênciasRESUMO
Evidence-based guidelines for monitoring patients with disorders in fatty acid oxidation (FAO) are lacking, and most protocols are based on expert statements. Here, we describe our protocol for Danish patients. Clinical monitoring is the most important measure and has the main aims of checking growth, development and diet and of bringing families to the clinic regularly to remind them of their child's risk and review how they cope and adjust, e.g. to an acute intercurrent illness. Most of these measures are simple and can be carried out during a routine out-patient visit; we seldom do more complicated assessments by a neuropsychologist, speech therapist, or physical and occupational therapists. Paraclinical measurements are not used for short-chain and medium-chain disorders; electrocardiography (including 24 h monitoring) and echocardiography are done for most patients with long-chain and carnitine transporter deficiencies. Eye examination is done in all, and liver ultrasonography in some patients with long-chain 3-hydroxyacyl-coenzyme A dehydrogenase/tri-functional protein (LCHAD/TFP) deficiencies. Biochemical follow-up includes determination of free carnitine and acylcarnitines. Free carnitine is measured to monitor carnitine supplementation in patients with multiple acyl-coenzyme A dehydrogenase deficiency (MADD) and carnitine transporter deficiency (CTD) and to follow metabolic control and disclose deficiency states in other FAO disorders. We are evaluating long-chain acylcarnitines in patients with long-chain disorders; so far there does not seem to be any clear-cut benefit in following these levels. An erythrocyte fatty acid profile is done in patients with long-chain disorders to test for essential fatty acid and docosahexanoic acid (DHA) deficiencies. The measurement of creatine kinase is helpful in long-chain disorders. Ongoing follow-up and education of the patient is important throughout life to prevent disease morbidity or death from metabolic crises.
Assuntos
Metabolismo Energético , Ácidos Graxos/metabolismo , Erros Inatos do Metabolismo Lipídico/diagnóstico , Mitocôndrias/enzimologia , Doenças Mitocondriais/diagnóstico , Adolescente , Adulto , Biomarcadores/sangue , Criança , Pré-Escolar , Continuidade da Assistência ao Paciente , Metabolismo Energético/genética , Genótipo , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Lactente , Recém-Nascido , Erros Inatos do Metabolismo Lipídico/enzimologia , Erros Inatos do Metabolismo Lipídico/genética , Erros Inatos do Metabolismo Lipídico/terapia , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Doenças Mitocondriais/terapia , Oxirredução , Educação de Pacientes como Assunto , Fenótipo , Valor Preditivo dos Testes , Prognóstico , Fatores de Tempo , Adulto JovemRESUMO
The accumulation of oxidatively modified proteins has been shown to be a characteristic feature of many neurodegenerative disorders and its regulation requires efficient proteolytic processing. One component of the mitochondrial proteolytic system is Lon, an ATP-dependent protease that has been shown to degrade oxidatively modified aconitase in vitro and may thus play a role in defending against the accumulation of oxidized matrix proteins in mitochondria. Using an assay system that allowed us to distinguish between basal and ATP-stimulated Lon protease activity, we have shown in isolated non-synaptic rat brain mitochondria that Lon protease is highly susceptible to oxidative inactivation by peroxynitrite (ONOO(-)). This susceptibility was more pronounced with regard to ATP-stimulated activity, which was inhibited by 75% in the presence of a bolus addition of 1mM ONOO(-), whereas basal unstimulated activity was inhibited by 45%. Treatment of mitochondria with a range of peroxynitrite concentrations (10-1000 microM) revealed that a decline in Lon protease activity preceded electron transport chain (ETC) dysfunction (complex I, II-III and IV) and that ATP-stimulated activity was approximately fivefold more sensitive than basal Lon protease activity. Furthermore, supplementation of mitochondrial matrix extracts with reduced glutathione, following ONOO(-) exposure, resulted in partial restoration of basal and ATP-stimulated activity, thus suggesting possible redox regulation of this enzyme complex. Taken together these findings suggest that Lon protease may be particularly vulnerable to inactivation in conditions associated with GSH depletion and elevated oxidative stress.
Assuntos
Encéfalo/enzimologia , Mitocôndrias/enzimologia , Doenças Mitocondriais/enzimologia , Estresse Oxidativo/fisiologia , Ácido Peroxinitroso/farmacologia , Protease La/metabolismo , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/farmacologia , Animais , Bioensaio , Encéfalo/fisiopatologia , Relação Dose-Resposta a Droga , Complexo de Proteínas da Cadeia de Transporte de Elétrons/efeitos dos fármacos , Complexo de Proteínas da Cadeia de Transporte de Elétrons/metabolismo , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/fisiologia , Glutationa/farmacologia , Masculino , Mitocôndrias/efeitos dos fármacos , Doenças Mitocondriais/fisiopatologia , Estresse Oxidativo/efeitos dos fármacos , Protease La/antagonistas & inibidores , Ratos , Ratos Wistar , Fatores de TempoRESUMO
The central nervous system plays a critical role in the normal control of arterial blood pressure and in its elevation in virtually all forms of hypertension. Mitochondrial dysfunction has been increasingly associated with the development of hypertension. Therefore, we examined whether mitochondrial dysfunction occurs in the brain in hypertension and characterized it at the molecular scale. Mitochondria from whole brain and brain stem from 12-week-old spontaneously hypertensive rats with elevated blood pressure (190+/-5 mm Hg) were compared against those from age-matched normotensive (134+/-7 mm Hg) Wistar Kyoto rats (n=4 in each group). Global differential analysis using 2D electrophoresis followed by tandem mass spectrometry-based protein identification suggested a downregulation of enzymes involved in cellular energetics in hypertension. Targeted differential analysis of mitochondrial respiratory complexes using the classical blue-native SDS-PAGE/Western method and a complementary combination of sucrose-gradient ultracentrifugation/tandem mass spectrometry revealed previously unknown assembly defects in complexes I, III, IV, and V in hypertension. Interestingly, targeted examination of the brain stem, a regulator of cardiovascular homeostasis and systemic blood pressure, further showed the occurrence of mitochondrial complex I dysfunction, elevated reactive oxygen species production, decreased ATP synthesis, and impaired respiration in hypertension. Our findings suggest that in already-hypertensive spontaneously hypertensive rats, the brain respiratory complexes exhibit previously unknown assembly defects. These defects impair the function of the mitochondrial respiratory chain. This mitochondrial dysfunction localizes to the brain stem and is, therefore, likely to contribute to the development, as well as to pathophysiological complications, of hypertension.
Assuntos
Encéfalo/enzimologia , Hipertensão/complicações , Hipertensão/fisiopatologia , Doenças Mitocondriais/etiologia , Complexos Multienzimáticos/genética , Processamento de Proteína Pós-Traducional , Animais , Sistema Cardiovascular/fisiopatologia , Doxiciclina/farmacologia , Eletroforese em Gel Bidimensional , Inibidores Enzimáticos/farmacologia , Homeostase , Hipertensão/enzimologia , Metaloendopeptidases/antagonistas & inibidores , Doenças Mitocondriais/enzimologia , Complexos Multienzimáticos/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Proteômica , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Espectrometria de Massas em TandemRESUMO
The cardiofaciocutaneous (CFC) syndrome is characterized by congenital heart defect, developmental delay, peculiar facial appearance with bitemporal constriction, prominent forehead, downslanting palpebral fissures, curly sparse hair and abnormalities of the skin. CFC syndrome phenotypically overlaps with Noonan and Costello syndromes. Mutations of several genes (PTPN11, HRAS, KRAS, BRAF, MEK1 and MEK2), involved in the mitogen-activated protein kinase (MAPK) pathway, have been identified in CFC-Costello-Noonan patients. Coenzyme Q10 (CoQ10), a lipophilic molecule present in all cell membranes, functions as an electron carrier in the mitochondrial respiratory chain, where it transports electrons from complexes I and II to complex III. CoQ10 deficiency is a rare treatable mitochondrial disorder with various neurological (cerebellar ataxia, myopathy, epilepsy, mental retardation) and extraneurological (cardiomyopathy, nephropathy) signs that are responsive to CoQ10 supplementation. We report the case of a 4-year-old girl who presented a CFC syndrome, confirmed by the presence of a pathogenic R257Q BRAF gene mutation, together with a muscular CoQ10 deficiency. Her psychomotor development was severely impaired, hindered by muscular hypotonia and ataxia, both improving remarkably after CoQ10 treatment. This case suggests that there is a functional connection between the MAPK pathway and the mitochondria. This could be through the phosphorylation of a nuclear receptor essential for CoQ10 biosynthesis. Another hypothesis is that K-Ras, one of the proteins composing the MAPK pathway, might be recruited into the mitochondria to promote apoptosis. This case highlights that CoQ10 might contribute to the pathogenesis of CFC syndrome.
Assuntos
Anormalidades Múltiplas , Anormalidades Craniofaciais/complicações , Cardiopatias Congênitas/complicações , Doenças Mitocondriais/complicações , Músculo Esquelético/enzimologia , Anormalidades da Pele/complicações , Ubiquinona/análogos & derivados , Anormalidades Múltiplas/enzimologia , Pré-Escolar , Coenzimas/deficiência , Coenzimas/uso terapêutico , Anormalidades Craniofaciais/enzimologia , Feminino , Cardiopatias Congênitas/enzimologia , Humanos , Sistema de Sinalização das MAP Quinases , Mitocôndrias/enzimologia , Doenças Mitocondriais/tratamento farmacológico , Doenças Mitocondriais/enzimologia , Anormalidades da Pele/enzimologia , Síndrome , Resultado do Tratamento , Ubiquinona/deficiência , Ubiquinona/uso terapêuticoRESUMO
This report presents a case of mitochondrial respiratory chain deficiency in a neonate with elevated plasma lactate, hypotonia, developmental delay, and dysmorphic features. The initial biochemical analyses of muscle tissue for mitochondrial function were normal. Additional testing on skin fibroblasts performed owing to a high clinical suspicion of a possible mitochondrial disorder indicated a deficiency of mitochondrial complex I. Western blotting of samples obtained both from muscle and fibroblast tissues also revealed an extensive defect in mitochondrial respiratory chain complex I, confirming the diagnosis. These observations underscore the fact that both enzymatic and immunological assays should be undertaken in alternate tissues when muscle biopsies are inconclusive in highly suspected cases.
Assuntos
Complexo I de Transporte de Elétrons/deficiência , Doenças Mitocondriais/diagnóstico , Músculo Esquelético/enzimologia , Acidose Láctica/diagnóstico , Acidose Láctica/enzimologia , Acidose Láctica/genética , Acidose Láctica/patologia , Atrofia , Biópsia , Western Blotting , Carnitina/análogos & derivados , Carnitina/sangue , Cistos do Sistema Nervoso Central/diagnóstico , Cistos do Sistema Nervoso Central/enzimologia , Cistos do Sistema Nervoso Central/genética , Cistos do Sistema Nervoso Central/patologia , Ventrículos Cerebrais/patologia , Análise Mutacional de DNA , Diagnóstico Diferencial , Complexo I de Transporte de Elétrons/genética , Feminino , Fibroblastos/enzimologia , Fibroblastos/patologia , Lobo Frontal/patologia , Humanos , Recém-Nascido , Imageamento por Ressonância Magnética , Doenças Mitocondriais/enzimologia , Doenças Mitocondriais/genética , Doenças Mitocondriais/patologia , Músculo Esquelético/patologia , Fibras Nervosas Mielinizadas/patologia , Mutação PuntualRESUMO
Rhizophora apiculata bark extract was tested for its free radical scavenging activity and protective role against mitochondrial dysfunction in naphthalene stressed rats. Lipid peroxidation activity was increased and activity of mitochondrial enzymes (cytochrome-c-oxidase, NADH-dehydrogenase, alpha-ketoglutarate dehydrogenase and succinate dehydrogenase) and glutathione was decreased in the liver and kidney of rats intoxicated with naphthalene when compared to control rats. Intraperitoneal administration of plant extract significantly reduced the lipid peroxidation, increased the activity of mitochondrial enzymes and increased glutathione to near control levels. These results suggest that the sulfated polysaccharides in R. apiculata play a protective role through their free radical scavenging properties.
Assuntos
Doenças Mitocondriais/tratamento farmacológico , Fitoterapia/métodos , Extratos Vegetais/farmacologia , Rhizophoraceae/química , Animais , Sequestradores de Radicais Livres/farmacologia , Glutationa/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Peróxidos Lipídicos/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Mitocôndrias/enzimologia , Doenças Mitocondriais/induzido quimicamente , Doenças Mitocondriais/enzimologia , Naftalenos , Casca de Planta/química , Ratos , Ratos WistarRESUMO
Mitochondria play a central role in heart metabolism and function. Administration of antineoplastic drug cyclophosphamide (CP) adversely affects the heart mitochondria which may result in cardiotoxicity. The present study is aimed at evaluating the role of lipoic acid (LA) in CP induced myocardial injury. Male albino rats of Wistar strain were used for the study. CP was administered as a single intraperitoneal injection (200 mg/kg BW). A decrease in the activities of TCA cycle enzymes such as succinate dehydrogenase, malate dehydrogenase and isocitrate dehydrogenase was noted in CP treated rats. Simultaneously there was a decrease in the activities of mitochondrial complexes of electron transport chain. Decrease in the activities of these enzymes suggests a loss in mitochondrial function and integrity. Ultrastuctural observations were also in agreement with the above abnormal changes. Loss of myofilaments and damage of mitochondrial cristae revealed the cytotoxic effect of CP. The supplementation of LA (25 mg/kg BW) restored the above abnormalities to near normalcy. The study brings out the importance of LA in improving the mitochondrial function in cardiac cells after CP administration.