RESUMO
Rett syndrome (RTT) is a neurodevelopmental disease that leads to intellectual deficit, motor disability, epilepsy and increased risk of sudden death. Although in up to 95% of cases this disease is caused by de novo loss-of-function mutations in the X-linked methyl-CpG binding protein 2 gene, it is a multisystem disease associated also with mitochondrial metabolic imbalance. In addition, the presence of long QT intervals (LQT) on the patients' electrocardiograms has been associated with the development of ventricular tachyarrhythmias and sudden death. In the attempt to shed light on the mechanism underlying heart failure in RTT, we investigated the contribution of the carnitine cycle to the onset of mitochondrial dysfunction in the cardiac tissues of two subgroups of RTT mice, namely Mecp2+/- NQTc and Mecp2+/- LQTc mice, that have a normal and an LQT interval, respectively. We found that carnitine palmitoyltransferase 1 A/B and carnitine acylcarnitine translocase were significantly upregulated at mRNA and protein level in the heart of Mecp2+/- mice. Moreover, the carnitine system was imbalanced in Mecp2+/- LQTc mice due to decreased carnitine acylcarnitine transferase expression. By causing accumulation of intramitochondrial acylcarnitines, this imbalance exacerbated incomplete fatty acid oxidation, which, in turn, could contribute to mitochondrial overload and sudden death.
Assuntos
Carnitina/metabolismo , Redes e Vias Metabólicas/genética , Síndrome de Rett/genética , Síndrome de Rett/metabolismo , Acetilcoenzima A/metabolismo , Animais , Carnitina/análogos & derivados , Modelos Animais de Doenças , Eletrocardiografia , Feminino , Genes Ligados ao Cromossomo X , Metabolismo dos Lipídeos , Metabolômica/métodos , Proteína 2 de Ligação a Metil-CpG/genética , Camundongos , Camundongos Knockout , Mitocôndrias/genética , Mitocôndrias/metabolismo , Mutação , Oxirredução , Fenótipo , Complexo Piruvato Desidrogenase/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Síndrome de Rett/diagnósticoRESUMO
Cancer cells are characterized by perturbations of their metabolic processes. Recent observations demonstrated that the fatty acid oxidation (FAO) pathway may represent an alternative carbon source for anabolic processes in different tumors, therefore appearing particularly promising for therapeutic purposes. Because the carnitine palmitoyl transferase 1a (CPT1a) is a protein that catalyzes the rate-limiting step of FAO, here we investigated the in vitro antileukemic activity of the novel CPT1a inhibitor ST1326 on leukemia cell lines and primary cells obtained from patients with hematologic malignancies. By real-time metabolic analysis, we documented that ST1326 inhibited FAO in leukemia cell lines associated with a dose- and time-dependent cell growth arrest, mitochondrial damage, and apoptosis induction. Data obtained on primary hematopoietic malignant cells confirmed the FAO inhibition and cytotoxic activity of ST1326, particularly on acute myeloid leukemia cells. These data suggest that leukemia treatment may be carried out by targeting metabolic processes.
Assuntos
Carnitina O-Palmitoiltransferase/antagonistas & inibidores , Carnitina/análogos & derivados , Sistemas de Liberação de Medicamentos , Ácidos Graxos/metabolismo , Leucemia Mieloide Aguda/tratamento farmacológico , Proteínas de Neoplasias/antagonistas & inibidores , Carnitina/farmacologia , Carnitina O-Palmitoiltransferase/metabolismo , Linhagem Celular Tumoral , Humanos , Leucemia Mieloide Aguda/enzimologia , Leucemia Mieloide Aguda/patologia , Proteínas de Neoplasias/metabolismo , Oxirredução/efeitos dos fármacosRESUMO
Oxaliplatin-based chemotherapy improves the outcomes of metastatic colorectal cancer patients. Its most significant and dose-limiting side effect is the development of a neuropathic syndrome. The mechanism of the neurotoxicity is unclear. The limited knowledge about differences existing between neurotoxic and antitumor effects hinders the discovery of effective and safe adjuvant therapies. In vitro, we suggested cell-specific activation apoptotic pathways in normal nervous cells (astrocytes) vs. colon-cancer cells (HT-29). In the present research we compared the apoptotic signals evoked by oxaliplatin in astrocytes and HT-29 analyzing the intrinsic and extrinsic apoptotic pathways. In astrocytes, oxaliplatin induced a mitochondrial derangement measured as cytosolic release of cytochrome C, increase in superoxide anion levels and decreased expression of the antiapoptotic protein Bcl-2. Caspase-8, a main initiator of the extrinsic process remained unaltered. On the contrary, in HT-29 oxaliplatin increased caspase-8 activity and Bid expression, thus activating the extrinsic apoptosis, while the Bcl-2 increased expression blocked the mitochondrial damage. Data suggest the preferred activation of the intrinsic apoptosis as oxaliplatin damage signaling in normal nervous cells. The extrinsic pathway prevails in tumor cells indicating a possible strategy for planning new molecules to treat oxaliplatin-dependent neurotoxicity without negatively influence chemotherapy.
Assuntos
Antineoplásicos/farmacologia , Apoptose , Astrócitos/efeitos dos fármacos , Neoplasias Colorretais/metabolismo , Compostos Organoplatínicos/farmacologia , Animais , Antineoplásicos/toxicidade , Astrócitos/metabolismo , Células Cultivadas , Células HT29 , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Compostos Organoplatínicos/toxicidade , Oxaliplatina , Células PC12 , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: The metabolic alterations of cancer cells represent an opportunity for developing selective antineoplastic treatments. We investigated the therapeutic potential of ST1326, an inhibitor of carnitine-palmitoyl transferase 1A (CPT1A), the rate-limiting enzyme for fatty acid (FA) import into mitochondria. METHODS: ST1326 was tested on in vitro and in vivo models of Burkitt's lymphoma, in which c-myc, which drives cellular demand for FA metabolism, is highly overexpressed. We performed assays to evaluate the effect of ST1326 on proliferation, FA oxidation, and FA mitochondrial channeling in Raji cells. The therapeutic efficacy of ST1326 was tested by treating Eµ-myc mice (control: n = 29; treatment: n = 24 per group), an established model of c-myc-mediated lymphomagenesis. Experiments were performed on spleen-derived c-myc-overexpressing B cells to clarify the role of c-myc in conferring sensitivity to ST1326. Survival was evaluated with Kaplan-Meier analyses. All statistical tests were two-sided. RESULTS: ST1326 blocked both long- and short-chain FA oxidation and showed a strong cytotoxic effect on Burkitt's lymphoma cells (on Raji cells at 72 hours: half maximal inhibitory concentration = 8.6 µM). ST1326 treatment induced massive cytoplasmic lipid accumulation, impairment of proper mitochondrial FA channeling, and reduced availability of cytosolic acetyl coenzyme A, a fundamental substrate for de novo lipogenesis. Moreover, treatment with ST1326 in Eµ-myc transgenic mice prevented tumor formation (P = .01), by selectively impairing the growth of spleen-derived primary B cells overexpressing c-myc (wild-type cells + ST1326 vs. Eµ-myc cells + ST1326: 99.75% vs. 57.5%, difference = 42.25, 95% confidence interval of difference = 14% to 70%; P = .01). CONCLUSIONS: Our data indicate that it is possible to tackle c-myc-driven tumorigenesis by altering lipid metabolism and exploiting the neoplastic cell addiction to FA oxidation.
Assuntos
Anticarcinógenos/farmacologia , Apoptose/efeitos dos fármacos , Linfoma de Burkitt/enzimologia , Linfoma de Burkitt/prevenção & controle , Carnitina O-Palmitoiltransferase/antagonistas & inibidores , Carnitina O-Palmitoiltransferase/metabolismo , Carnitina/análogos & derivados , Metabolismo dos Lipídeos/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-myc/metabolismo , Animais , Linfócitos B/efeitos dos fármacos , Linfócitos B/metabolismo , Western Blotting , Linfoma de Burkitt/metabolismo , Linfoma de Burkitt/patologia , Carnitina/farmacologia , Carnitina Aciltransferases/antagonistas & inibidores , Carnitina Aciltransferases/metabolismo , Linhagem Celular Tumoral , Ácidos Graxos/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Estimativa de Kaplan-Meier , Camundongos , Oxirredução , Proteínas Proto-Oncogênicas c-myc/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Resultado do Tratamento , Regulação para CimaRESUMO
Maternal-fetal HIV-1 transmission can be prevented by administration of AZT, alone or in combination with other antiretroviral drugs to pregnant HIV-1-infected women and their newborns. In spite of the benefits deriving from this life-saving prophylactic therapy, there is still considerable uncertainty on the potential long-term adverse effects of antiretroviral drugs on exposed children. Clinical and experimental studies have consistently shown the occurrence of mitochondrial dysfunction and increased oxidative stress following prenatal treatment with antiretroviral drugs, and clinical evidence suggests that the developing brain is one of the targets of the toxic action of these compounds possibly resulting in behavioral problems. We intended to verify the effects on brain and behavior of mice exposed during gestation to AZT, the backbone of antiretroviral therapy during human pregnancy. We hypothesized that glutamate, a neurotransmitter involved in excitotoxicity and behavioral plasticity, could be one of the major actors in AZT-induced neurochemical and behavioral alterations. We also assessed the antioxidant and neuroprotective effect of L-acetylcarnitine, a compound that improves mitochondrial function and is successfully used to treat antiretroviral-induced polyneuropathy in HIV-1 patients. We found that transplacental exposure to AZT given per os to pregnant mice from day 10 of pregnancy to delivery impaired in the adult offspring spatial learning and memory, enhanced corticosterone release in response to acute stress, increased brain oxidative stress also at birth and markedly reduced expression of mGluR1 and mGluR5 subtypes and GluR1 subunit of AMPA receptors in the hippocampus. Notably, administration during the entire pregnancy of L-acetylcarnitine was effective in preventing/ameliorating the neurochemical, neuroendocrine and behavioral adverse effects induced by AZT in the offspring. The present preclinical findings provide a mechanistic hypothesis for the neurobehavioral effects of AZT and strongly suggest that preventive administration of L-acetylcarnitine might be effective in reducing the neurological side-effects of antiretroviral therapy in fetus/newborn.
Assuntos
Acetilcarnitina/farmacologia , Fármacos Anti-HIV/efeitos adversos , Comportamento Animal/efeitos dos fármacos , Troca Materno-Fetal , Fármacos Neuroprotetores/farmacologia , Zidovudina/efeitos adversos , Animais , Fármacos Anti-HIV/administração & dosagem , Peso ao Nascer/efeitos dos fármacos , Feminino , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Memória/efeitos dos fármacos , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Gravidez , Estresse Fisiológico/efeitos dos fármacos , Zidovudina/administração & dosagemRESUMO
INTRODUCTION: L-Acetylcarnitine (LAC), the acetyl ester of carnitine naturally present in the central nervous system and involved in several neural pathways, has been demonstrated to be active in various animal experimental models resembling some features of human depression. The aim of the study is to verify whether LAC can have an antidepressant action in a population of elderly patients with dysthymic disorder in comparison with a traditional antidepressant such as fluoxetine. METHODS: Multicentric, double-blind, double-dummy, controlled, randomized study based on a observation period of 7 weeks. 80 patients with DSM-IV diagnosis of dysthymic disorder were enrolled in the study and subdivided into 2 groups. Group A patients received LAC plus placebo; group B patients received fluoxetine 20 mg/die plus placebo. Clinical assessment was performed through several psychometric scales at 6 different moments. RESULTS: Group A patients showed a statistically significant improvement in the following scales: HAM-D, HAM-A, BDI and Touluse Pieron Test. Comparison between the two groups, A and B, generally showed very similar clinical progression. DISCUSSION: The results obtained with LAC and fluoxetine were equivalent. As the subjects in this study were of senile age, it is possible to hypothesize that the LAC positive effect on mood could be associated with improvement in subjective cognitive symptomatology. The difference in the latency time of clinical response (1 week of LAC treatment, compared with the 2 weeks' latency time with fluoxetine) suggests the existence of different mechanisms of action possibly in relation to the activation of rapid support processes of neuronal activity.
Assuntos
Acetilcarnitina/uso terapêutico , Envelhecimento , Antidepressivos/uso terapêutico , Transtorno Distímico/tratamento farmacológico , Acetilcarnitina/efeitos adversos , Idoso , Idoso de 80 Anos ou mais , Antidepressivos/efeitos adversos , Disfunção Cognitiva/tratamento farmacológico , Disfunção Cognitiva/fisiopatologia , Disfunção Cognitiva/psicologia , Manual Diagnóstico e Estatístico de Transtornos Mentais , Método Duplo-Cego , Transtorno Distímico/etiologia , Transtorno Distímico/fisiopatologia , Feminino , Fluoxetina/efeitos adversos , Fluoxetina/uso terapêutico , Humanos , Masculino , Nootrópicos/efeitos adversos , Nootrópicos/uso terapêutico , Escalas de Graduação Psiquiátrica , Índice de Gravidade de Doença , Equivalência Terapêutica , Fatores de TempoRESUMO
Rett syndrome (RTT) is a devastating neurodevelopmental disorder affecting 1 in 10,000 girls. Approximately 90% of cases are caused by spontaneous mutations in the X-linked gene encoding methyl-CpG-binding protein 2 (MeCP2). Girls with RTT suffer from severe motor, respiratory, cognitive and social abnormalities attributed to early deficits in synaptic connectivity which manifest in the adult as a myriad of physiological and anatomical abnormalities including, but not limited to, dimished dendritic complexity. Supplementation with acetyl-L-carnitine (ALC), an acetyl group donor, ameliorates motor and cognitive deficits in other disease models through a variety of mechanisms including altering patterns of histone acetylation resulting in changes in gene expression, and stimulating biosynthetic pathways such as acetylcholine. We hypothesized ALC treatment during critical periods in cortical development would promote normal synaptic maturation, and continuing treatment would improve behavioral deficits in the Mecp2(1lox) mouse model of RTT. In this study, wildtype and Mecp2(1lox) mutant mice received daily injections of ALC from birth until death (postnatal day 47). General health, motor, respiratory, and cognitive functions were assessed at several time points during symptom progression. ALC improved weight gain, grip strength, activity levels, prevented metabolic abnormalities and modestly improved cognitive function in Mecp2 null mice early in the course of treatment, but did not significantly improve motor or cognitive functions assessed later in life. ALC treatment from birth was associated with an almost complete rescue of hippocampal dendritic morphology abnormalities with no discernable side effects in the mutant mice. Therefore, ALC appears to be a promising therapeutic approach to treating early RTT symptoms and may be useful in combination with other therapies.
Assuntos
Acetilcarnitina/uso terapêutico , Comportamento Animal , Dendritos/patologia , Síndrome de Rett/tratamento farmacológico , Síndrome de Rett/patologia , Acetilcarnitina/sangue , Acetilcarnitina/farmacologia , Animais , Animais Recém-Nascidos , Comportamento Animal/efeitos dos fármacos , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Cognição/efeitos dos fármacos , Dendritos/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Heterozigoto , Hipocampo/efeitos dos fármacos , Hipocampo/patologia , Hipocampo/fisiopatologia , Masculino , Proteína 2 de Ligação a Metil-CpG/deficiência , Proteína 2 de Ligação a Metil-CpG/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Atividade Motora/efeitos dos fármacos , Fator de Crescimento Neural/metabolismo , Síndrome de Rett/sangue , Síndrome de Rett/fisiopatologiaRESUMO
RATIONALE: Depression may be associated with altered plasticity of the nervous system. The importance of neurotrophic factor levels is strongly suggested, and the neuronal-related family is extensively studied with respect to glial-derived one. OBJECTIVES: Aimed to contribute to the study of nervous plasticity modulation as therapeutical target in mood disorders, the role of the glial-derived factor artemin (ARTN) in depression and in the pharmacodynamics of the antidepressant and trophic compound acetyl-L: -carnitine (ALCAR) was evaluated. METHODS: Male mice were treated with 100 mg kg(-1) ALCAR daily for 7 days; 0.6 µg/mouse ARTN was acutely injected intracerebroventricularly. Gene knockdown of ARTN and GDNF family receptor alpha (GFRalpha3) was obtained by oligonucleotide antisense strategy. The forced swimming test was performed to evaluate antidepressant-like effects. RESULTS: Repeated ALCAR administration increased ARTN levels in spinal cord, hippocampus, and prefrontal cortex. No modulatory effect was detected on BDNF and glial cell line-derived neutrotrophic factor (GDNF). ARTN, 30 min after administration, showed a dose-dependent antidepressant-like effect. ALCAR needed a 7-day treatment to reach a comparable effect; nevertheless, both substances were able to induce a phosphorylation of the GDNF family receptor Ret. A decrease of the free ARTN level by a specific ARTN antibody impaired the antidepressant-like effect of acute ARTN and repeated ALCAR. Gene knockdown of ARTN or, alternatively, of its receptor GFRalpha3 fully prevented ALCAR effectiveness. CONCLUSIONS: A mechanism for the antidepressant property of ALCAR is proposed, and the novelty of the possible role of ARTN in depression is suggested.
Assuntos
Acetilcarnitina/farmacologia , Antidepressivos/farmacologia , Depressão/tratamento farmacológico , Proteínas do Tecido Nervoso/metabolismo , Acetilcarnitina/administração & dosagem , Animais , Antidepressivos/administração & dosagem , Depressão/fisiopatologia , Modelos Animais de Doenças , Técnicas de Silenciamento de Genes , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Hipocampo/metabolismo , Injeções Intraventriculares , Masculino , Camundongos , Proteínas do Tecido Nervoso/administração & dosagem , Proteínas do Tecido Nervoso/genética , Oligonucleotídeos Antissenso/administração & dosagem , Córtex Pré-Frontal/metabolismo , Medula Espinal/metabolismo , NataçãoRESUMO
Several pathologies related to nervous tissue alterations are characterized by a chronic pain syndrome defined by persistent or paroxysmal pain independent or dependent on a stimulus. Pathophysiological mechanisms related to neuropathic disease are associated with mitochondrial dysfunctions that lead to an activation of the apoptotic cascade. In a model of peripheral neuropathy obtained by the loose ligation of the rat sciatic nerve, acetyl-L-Carnitine (ALCAR; 100 mg/kg intraperitoneally [i.p.] twice daily for 14 days) was able to reduce hyperalgesia and apoptosis. In the present study, different mechanisms for the analgesic and the antineuropathic effect of ALCAR are described. The muscarinic blocker atropine (5 mg/kg i.p.) injected simultaneously with ALCAR did not antagonize the ALCAR antihyperalgesic effect on the paw-pressure test but significantly reduced the analgesic effect of ALCAR. Conversely, the antineuropathic effect of ALCAR was prevented by cotreatment with the nicotinic antagonist mecamylamine (2 mg/kg i.p. twice daily for 14 days). A pharmacological silencing of the nicotinic receptors significantly reduced the X-linked inhibitor of apoptosis protein-related protective effect of ALCAR on the apoptosis induced by ligation of the sciatic nerve. Taken together, these data highlight the relevance of nicotinic modulation in neuropathy treatment.
Assuntos
Acetilcarnitina/uso terapêutico , Apoptose/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Receptores Nicotínicos/metabolismo , Ciática , Animais , Atropina/farmacologia , Caspase 3/metabolismo , Citocromos c/metabolismo , Modelos Animais de Doenças , Marcação In Situ das Extremidades Cortadas/métodos , Masculino , Mecamilamina/farmacologia , Antagonistas Muscarínicos/farmacologia , Antagonistas Nicotínicos/farmacologia , Medição da Dor/efeitos dos fármacos , Medição da Dor/métodos , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/metabolismo , Ratos , Ratos Sprague-Dawley , Ciática/tratamento farmacológico , Ciática/patologia , Ciática/fisiopatologia , Fatores de Tempo , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/metabolismoRESUMO
INTRODUCTION: This study was designed to determine the short-term effect of acetyl-l-carnitine (ALC) on symptoms of withdrawal in opiate-dependent subjects and animals and, in particular, on pain, given the efficacy of ALC in other typologies of pain. The study consists of 2 branches: a clinical study and a preclinical one, both with a randomized placebo-controlled design. METHODS: Thirty subjects meeting clinical criteria for methadone dependence were consecutively recruited and treated with ALC 2 g/d or placebo for a 3-week detoxification period. Withdrawal symptoms and pain were evaluated through the Short Opiate Withdrawal Syndrome scale, and the Huskisson's analogue scale for pain. In the preclinical study, mice previously received a pretreatment (saline solution or morphine), and subsequently, each group was randomly divided in 4 subgroups that received a treatment of saline, methadone, ALC, or amitriptyline, respectively. Hot plate test and Writhing test were used to evaluate pain intensity. RESULTS: Average Short Opiate Withdrawal Syndrome total scores during the first 5 days of treatment resulted significantly higher in controls than in the ALC group (P < 0.05). Pain scores in the Huskisson's analogue scale were considerably lower in the group of patients taking ALC than in the control group after 1 week of ALC treatment until the end of the study. Results of the preclinical study show that the administration of methadone for 7 days in morphine-tolerant mice did not produce any modification of the pain threshold. By contrast, the 7-day coadministration of methadone and ALC in morphine-tolerant mice induced an analgesic effect evaluated 3 hours after the last injection. DISCUSSION: Acetyl-L-carnitine acted as an effective antihyperalgesic agent for relieving opiate-withdrawal hyperalgesia in animals and displayed clinical efficacy on other withdrawal symptoms such as muscular tension, muscular cramps, and insomnia. Considering its tolerability, the excellent side effect profile, the absence of significant interactions, and the lack of abuse potential, ALC can be considered as a useful pharmacological adjunct in the treatment of opiate withdrawal.
Assuntos
Acetilcarnitina/uso terapêutico , Analgésicos Opioides/efeitos adversos , Metadona/efeitos adversos , Nootrópicos/uso terapêutico , Dor/tratamento farmacológico , Dor/etiologia , Síndrome de Abstinência a Substâncias/complicações , Adulto , Amitriptilina/uso terapêutico , Analgésicos não Narcóticos/uso terapêutico , Analgésicos Opioides/administração & dosagem , Animais , Modelos Animais de Doenças , Esquema de Medicação , Feminino , Humanos , Masculino , Metadona/administração & dosagem , Camundongos , Morfina/administração & dosagem , Transtornos Relacionados ao Uso de Opioides/complicações , Transtornos Relacionados ao Uso de Opioides/tratamento farmacológico , Medição da Dor , Fatores de Tempo , Adulto JovemRESUMO
Since its definition Alzheimer's disease has been at the centre of consideration for neurologists, psychiatrists, and pathologists. With John P. Blass it has been disclosed a different approach Alzheimer's disease neurodegeneration understanding not only by the means of neurochemistry but also biochemistry opening new scenarios in the direction of a metabolic system degeneration. Nowadays, the understanding of the role of cholesterol, insulin, and adipokines among the others in Alzheimer's disease etiopathogenesis is clarifying approaches valuable not only in preventing the disease but also for its therapy.
Assuntos
Doença de Alzheimer/história , Doença de Alzheimer/metabolismo , Química Encefálica/fisiologia , Doenças Metabólicas/metabolismo , Adiponectina/metabolismo , Doença de Alzheimer/psicologia , Apolipoproteínas E/metabolismo , Colesterol/metabolismo , Feminino , Grelina , História do Século XX , Humanos , Insulina/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Leptina/metabolismo , Pessoa de Meia-Idade , Hormônios Peptídicos/metabolismoRESUMO
Immunohistology of lymphoreticular tissues of a fatal case of organic cation transporter 2 deficiency revealed inhibited proliferation with increased apoptosis in the germinal centers, resulting in "burned out" follicles. This is indicative of impaired antigen driven B cell affinity maturation. Defective humoral immune response might explain the recurrent infections in untreated organic cation transporter 2 deficiency.
Assuntos
Linfócitos B/patologia , Crescimento Celular , Tecido Linfoide/patologia , Proteínas de Transporte de Cátions Orgânicos/deficiência , Infecções Respiratórias/patologia , Proliferação de Células , Pré-Escolar , Progressão da Doença , Humanos , Lactente , Tecido Linfoide/metabolismo , Masculino , Infecções Respiratórias/imunologia , Infecções Respiratórias/metabolismo , Membro 5 da Família 22 de Carreadores de SolutoRESUMO
The neuronal death after ischemia is closely linked to the essential role of mitochondrial metabolism. Inhibition of mitochondrial respiratory chain reduces ATP generation leading to a dysregulation of ion metabolism. Acetyl-L-carnitine (ALC) influences the maintenance of key mitochondrial proteins for maximum energy production and it may play a neuroprotective role in some pathological conditions. In this study we have analyzed ALC-mediated neuroprotection on an in vitro model of brain ischemia. Field potential recordings were obtained from a rat corticostriatal slice preparation. In vitro ischemia (oxygen and glucose deprivation) was delivered by switching to a solution in which glucose was omitted and oxygen was replaced with N2. Ten minutes of in vitro ischemia caused an irreversible loss of the field potential amplitude. Pretreatment with ALC produced a progressive and dose-dependent recovery of the field potential amplitude following in vitro ischemia. The neuroprotective effect of ALC was stereospecific since the pretreatment with two different carnitine-related compounds did not cause neuroprotection. The choline transporter inhibitor hemicholinium-3 blocked the neuroprotective effect of ALC. ALC-mediated neuroprotection was also prevented either by the non-selective muscarinic antagonist scopolamine, or by the putative M2-like receptor antagonist methoctramine. Conversely, the effect of ALC was not altered by the M1-like receptor antagonist pirenzepine. These findings show that ALC exert a neuroprotective action against in vitro ischemia. This neuroprotective effect requires the activity of choline uptake system and the activation of M2 muscarinic receptors.
Assuntos
Acetilcarnitina/uso terapêutico , Acetilcolina/fisiologia , Isquemia/prevenção & controle , Neurônios/efeitos dos fármacos , Nootrópicos/uso terapêutico , Córtex Visual/patologia , Acetilcarnitina/análogos & derivados , Animais , Colinérgicos/farmacologia , Diaminas/farmacologia , Relação Dose-Resposta a Droga , Interações Medicamentosas , Potenciais Evocados/efeitos dos fármacos , Potenciais Evocados/fisiologia , Potenciais Evocados/efeitos da radiação , Hemicolínio 3/farmacologia , Técnicas In Vitro , Antagonistas Muscarínicos/farmacologia , Parassimpatolíticos/farmacologia , Pirenzepina/farmacologia , Ratos , Ratos Wistar , Escopolamina/farmacologiaRESUMO
Literature data indicate L-carnitine (LC), a trans-mitochondrial carrier of acetyl and long chain groups, as an agent possessing protective effects against oxidative stress in mammalian cells. However, the major factor involved in the protective mechanism is not known. The protection activity exerted by this agent against reactive oxygen species induced by hydrogen peroxide (H2O2) and t-butylhydroperoxide (t-butyl-OOH) treatment in isolated human peripheral blood lymphocytes (PBLs) has been studied. Human lymphocytes cells were isolated and pre-incubated with 5 mM LC before H2O2 (100 microM) and t-butyl-OOH (400 microM) treatment. The protective effect of LC on treated PBLs was measured by single cell gel electrophoresis and the analysis of chromosomal aberrations. Results show that lc treated cells exhibited a significant decrease in the number of oxidative induced single-strand breaks and chromosomal aberrations.
Assuntos
Carnitina/farmacologia , Aberrações Cromossômicas/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Oxidantes/farmacologia , Substâncias Protetoras/farmacologia , Complexo Vitamínico B/farmacologia , Ensaio Cometa , Humanos , Peróxido de Hidrogênio/farmacologia , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , terc-Butil Hidroperóxido/farmacologiaRESUMO
BACKGROUND: Increased oxidative stress is associated with all cardiovascular risk factors and reactive oxygen species appear to be the principal mediators of cardiomyocite dysfunction in various cardiovascular diseases. Carnitine has been shown to be effective in pathologic conditions characterized by increased oxidative stress and an antioxidant effect of L-carnitine and its derivatives has been described but the specific mechanism is unclear. METHODS: We evaluated in human endothelial cells in culture the effect of L-carnitine (C), acetyl-L-carnitine (AC) and propionyl-L-carnitine (PC) on gene and protein expression (RT-PCR and Western blot) of oxidative stress related proteins heme oxygenase-1 (HO-1) and of endothelial NO synthase (ecNOS) in absence and presence of oxidative stress induced by H2O2. RESULTS: HO-1 as well as ecNOS gene and protein expression significantly increased upon Carnitines incubation. Induction of oxidative stress increased HO-1 gene expression compared to basal condition (0.62+/-0.02 densitometric units vs. 0.48+/-0.05, p<0.01) while decreased ecNOS gene expression (0.75+/-0.04 vs. 0.40+/-0.08, p<0.001). These results were paralleled by similar results at protein level. Coincubation of C (0.5-1.0-2.0 mM), AC (0.1-0.2-0.4 mM) and PC (0.05-0.1-0.2 mM) with H2O2 further increased HO-1 gene expression and not only normalized vs. H2O2 but even increased vs. basal ecNOS mRNA. HO-1 and ecNOS gene expression was also paralleled at protein level by coincubation with C, AC and PC of cells exposed to oxidative stress. CONCLUSION: This is the first report that has utilized a molecular biological approach to demonstrate a direct stimulatory effect of Carnitines on gene and protein expression of the oxidative stress related markers HO-1 and ecNOS. As HO-1 and NO are known as antioxidant, antiproliferative and anti-inflammatory, their increased expression would be expected to protect from oxidative stress related cardiovascular risk factors and myocardial damage, therefore adding this effect to the multiple pathways involved in the effects of carnitines.
Assuntos
Doenças Cardiovasculares/complicações , Carnitina/farmacologia , Endotélio/lesões , Estresse Oxidativo/fisiologia , Apoptose , Doenças Cardiovasculares/fisiopatologia , Técnicas de Cultura de Células , Células Endoteliais/efeitos dos fármacos , Endotélio/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Heme Oxigenase-1/efeitos dos fármacos , Heme Oxigenase-1/genética , Humanos , Óxido Nítrico Sintase/efeitos dos fármacos , Óxido Nítrico Sintase/genética , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio , Cordão UmbilicalRESUMO
Acetyl-L-carnitine (ALC) is a molecule widely present in the central nervous system (CNS) formed by the reversible acetylation of carnitine. It acts by stimulating energy metabolism. Reported neurobiological effects of this substance include modulation of brain energy and phospholipid metabolism; cellular macromolecules (including neurotrophic factors and neurohormones); synaptic transmission of multiple neurotransmitters. ALC is of considerable interest for its clinical application in Alzheimer's disease and in the treatment of painful neuropathies. There are experimental data that it affects attention and antagonizes deterioration of ability to learn, improving long-term memory. Moreover, ALC influences nonassociative learning of sensitization type in Hirudo medicinalis. These findings are suggesting that ALC might exert its effects by means of new protein synthesis. ALC or saline solution was injected intraperitoneally each day for 21 days in rats. Poly(A)+ RNAs were isolated from control and treated rat brain. Suppression subtractive hybridisation (SSH) method was applied for the generation of subtracted cDNA libraries and the subsequent identification of differentially expressed transcripts after treatments. The technique generates an equalized representation of differentially expressed genes irrespective of their relative abundance, and it is based on the construction of forward and reverse cDNA libraries that allow the identification of the genes that are regulated or switched off/on after ALC treatment. We identified two modulated genes, the isoform gamma of 14-3-3 protein and a precursor of ATP synthase lipid-binding protein, and one gene switched on by the treatment, the heat shock protein hsp72.
Assuntos
Acetilcarnitina/farmacologia , Encéfalo/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas do Tecido Nervoso/genética , Proteínas 14-3-3/biossíntese , Proteínas 14-3-3/genética , Acetilcarnitina/metabolismo , Animais , Encéfalo/metabolismo , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/genética , Biblioteca Gênica , Proteínas de Choque Térmico HSP72 , Proteínas de Choque Térmico/biossíntese , Proteínas de Choque Térmico/genética , Masculino , ATPases Mitocondriais Próton-Translocadoras/biossíntese , ATPases Mitocondriais Próton-Translocadoras/genética , Fatores de Crescimento Neural/metabolismo , Fatores de Crescimento Neural/farmacologia , Proteínas do Tecido Nervoso/biossíntese , Hibridização de Ácido Nucleico/métodos , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genéticaRESUMO
Dietary polyunsaturated fatty acids (PUFA) have effects on diverse physiological processes impacting normal health and chronic diseases, such as the regulation of plasma lipid levels, cardiovascular and immune function, insulin action and neuronal development and visual function. Ingestion of PUFA will lead to their distribution to virtually every cell in the body with effects on membrane composition and function, eicosanoid synthesis, cellular signaling and regulation of gene expression. Cell specific lipid metabolism, as well as the expression of fatty acid-regulated transcription factors, likely play an important role in determining how cells respond to changes in PUFA composition. This review will focus on recent advances on the essentiality of these molecules and on their interplay in cell physiology, leading to new perspective in different therapeutic fields.
Assuntos
Gorduras Insaturadas na Dieta/administração & dosagem , Gorduras na Dieta/administração & dosagem , Ácidos Graxos Insaturados/fisiologia , Regulação da Expressão Gênica , Animais , Doença Crônica , Ácidos Graxos Insaturados/metabolismo , Humanos , Estado Nutricional , Transcrição GênicaRESUMO
The cellular events involved in acetyl-L-carnitine (ALCAR) analgesia were investigated in the mouse hot plate test. I.c.v. pretreatment with aODNs against the alpha subunit of G(q) and G(11) proteins prevented the analgesia induced by ALCAR (100 mg kg(-1) s.c. twice daily for 7 days). Administration of the phospholipase C (PLC) inhibitors U-73122 and neomycin, as well as the injection of an aODN complementary to the sequence of PLCbeta(1), antagonized the increase of the pain threshold induced by ALCAR. Pretreatment with U-73343, an analogue of U-73112 inactive on PLC, did not modify ALCAR analgesic effect. In mice undergoing treatment with LiCl, which impairs phosphatidylinositol synthesis, or pretreatment with TMB-8, a blocker of Ca(++) release from intracellular stores, the antinociception induced by ALCAR was dose-dependently antagonized. I.c.v. treatment with heparin, an IP(3) receptor antagonist, prevented the increase of pain threshold induced by the investigated compound, analgesia that was restored by co-administration of D-myo-inositol. On the other hand, i.c.v. pretreatment with the selective protein kinase C (PKC) inhibitors calphostin C and cheleritryne, resulted in a dose-dependent potentiation of ALCAR antinociception. The administration of PKC activators, such as PMA and PDBu, dose-dependently prevented the ALCAR-induced increase of pain threshold. Neither aODNs nor pharmacological treatments produced any behavioral impairment of mice as revealed by the rota-rod and hole board tests. These results indicate that central ALCAR analgesia in mice requires the activation of the PLC-IP(3) pathway. By contrast, the simultaneous activation of PKC may represent a pathway of negative modulation of ALCAR antinociception.
Assuntos
Acetilcarnitina/farmacologia , Analgésicos , Inositol 1,4,5-Trifosfato/fisiologia , Nootrópicos/farmacologia , Fosfolipases Tipo C/fisiologia , Acetilcarnitina/antagonistas & inibidores , Analgésicos/antagonistas & inibidores , Animais , Comportamento Animal/efeitos dos fármacos , Diglicerídeos/farmacologia , Sinergismo Farmacológico , Ativação Enzimática/efeitos dos fármacos , Temperatura Alta , Injeções Intraventriculares , Cloreto de Lítio/farmacologia , Masculino , Camundongos , Nootrópicos/antagonistas & inibidores , Oligonucleotídeos Antissenso/farmacologia , Medição da Dor/efeitos dos fármacos , Equilíbrio Postural , Proteína Quinase C/fisiologia , Tempo de Reação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
The analgesic activity of acetyl-L-carnitine (ALCAR) in neuropathic pain is well established. By contrast, its potential efficacy in the relief of acute pain has not been reported. The antinociceptive effect of ALCAR was, therefore, examined in the mouse hot-plate and abdominal constriction tests, and in the rat paw-pressure test. ALCAR (100 mg kg(-1) s.c. twice daily for seven days) produced an increase of the pain threshold in both mice and rats. ALCAR was also able to reverse hyperalgesia induced by kainic acid and NMDA administration in the mouse hot-plate test. The antinociception produced by ALCAR was prevented by the unselective muscarinic antagonist atropine, the M(1) selective antagonists pirenzepine and S-(-)-ET126, and by the choline uptake inhibitor hemicholinium-3 (HC-3). By contrast the analgesic effect of ALCAR was not prevented by the opioid antagonist naloxone, the GABA(B) antagonist CGP 35348, the monoamine synthesis inhibitor (alpha)-methyl-p-tyrosine, and the Gi-protein inactivator pertussis toxin. Moreover, ALCAR antinociception was abolished by pretreament with an antisense oligonucleotide (aODN) against the M(1) receptor subtype, administered at the dose of 2 nmol per single i.c.v injection. On the basis of the above data, it can be postulated that ALCAR exerted an antinociceptive effect mediated by a central indirect cholinergic mechanism. In the antinociceptive dose-range, ALCAR did not impair mouse performance evaluated by the rota-rod and hole-board tests.