Implicación de la mitocondria y del complejo NLRP3-inflamasoma en la fisiopatología de la fibromialgia

La fibromialgia (FM) es un síndrome de dolor crónico generalizado de alta prevalencia mundial que supone un verdadero enigma clínico dado su heterogeneidad y su desconocida etiología. Su fisiopatología ha sido ampliamente investigada y se han propuesto varias hipótesis biológicas entre las cuales, la inflamación, la disfunción mitocondrial y el estrés oxidativo han mostrado una gran implicación. Nuestro estudio ha profundizado en el papel de la disfunción mitocondrial mediante el uso de un modelo in vitro utilizado para el estudio de las patologías mitocondriales. Para ello, hemos usado fibroblastos de piel mediante los cuales, hemos descrito una deficiencia de la actividad de la cadena respiratoria mitocondrial, una deficiencia de CoQ10, así como un incremento del daño oxidativo. Además, esta disfunción mitocondrial provocada la activación de un complejo multiproteico denominado, complejo NLRP3-inflamasoma el cual, es responsable se la inflamación sistémica. Por otro lado, muchos de los síntomas asociados con las enfermedades mitocondriales, como intolerancia al ejercicio, fatiga, miopatía están presentes en muchos pacientes con FM que presentan disfunción mitocondrial. En este estudio, mostramos una mutación en el gen citocromo b del ADN mitocondrial (ADNmt) en una familia con FM y la implicación del complejo NLRP3-inflamasoma en varias mutaciones mitocondriales. Tras la secuenciación de ADNmt de varios pacientes con FM, encontramos una mutación mitocondrial homoplásmica m.15804T > C en el gen mtCYB en un paciente transmitida por vía materna, que aparece en todos miembros de la familia con el diagnóstico de FM. La mutación estaba presente en varios tejidos de la paciente. Los fibroblastos de piel de la paciente mostraron disfunción mitocondrial y estrés oxidativo asociado a la activación del complejo NLRP3-inflamasoma así como una mejora del metabolismo celular tras la inhibición del complejo inflamasoma o la suplementación con CoQ10. Los cíbridos trans-mitocondriales que portaban la mutación m.15804T> C mostraron las alteraciones fisiopatológicas observadas en los fibroblastos de la paciente con FM. Interesantemente, la activación del complejo NLRP3-inflamasoma también se observó en líneas de fibroblastos derivados de pacientes con otras enfermedades mitocondriales como Miopatía mitocondrial, encefalopatía, acidosis láctica y accidentes cerebrovasculares (MELAS), Epilepsia mioclónica con fibras rojas rasgadas (MERRF) y Neuropatía óptica hereditaria de Leber (LHON). En este trabajo mostramos por primera vez una mutación en el ADNmt responsable de la disfunción mitocondrial, el estrés oxidativo y la activación inflamasoma asociado con una familia con diagnóstico de FM. Sobre la base de nuestros datos, se propone el análisis de secuencias del ADNmt en pacientes con FM con evidencias de herencia materna como una posible herramienta de diagnóstico. Además, la activación NLRP3-inflamasoma como consecuencia de la disfunción mitocondrial podría estar implicada en la fisiopatología de las enfermedades mitocondriales

Mitochondrial dysfunction promoted by Porphyromonas gingivalis lipopolysaccharide as a possible link between cardiovascular disease and periodontitis

Oxidative stress is one of the factors that could explain the pathophysiological mechanism of inflammatory conditions that occur in cardiovascular disease (CVD) and periodontitis. Such inflammatory response is often evoked by specific bacteria, as the lipopolysaccharide (LPS) of Porphyromonas gingivalis is a key factor in this process. The aim of this research was to study the role of mitochondrial dysfunction in peripheral blood mononuclear cells (PBMCs) from periodontitis patients and to evaluate the influence of LPS on fibroblasts to better understand the pathophysiology of periodontitis and its relationship with CVD. PBMCs from patients showed lower CoQ10 levels and citrate synthase activity, together with high levels of ROS production. LPS-treated fibroblasts provoked increased oxidative stress and mitochondrial dysfunction by a decrease in mitochondrial protein expression, mitochondrial mass, and mitochondrial membrane potential. Our study supports the hypothesis that LPS-mediated mitochondrial dysfunction could be at the origin of oxidative stress in periodontal patients. Abnormal PBMC performance may promote oxidative stress and alter cytokine homeostasis. In conclusion, mitochondrial dysfunction could represent a possible link to understanding the interrelationships between two prominent inflammatory diseases: periodontitis and CV

Current experience in testing mitochondrial nutrients in disorders featuring oxidative stress and mitochondrial dysfunction: rational design of chemoprevention trials

An extensive number of pathologies are associated with mitochondrial dysfunction (MDF) and oxidative stress (OS). Thus, mitochondrial cofactors termed “mitochondrial nutrients” (MN), such as α-lipoic acid (ALA), Coenzyme Q10 (CoQ10), and l-carnitine (CARN) (or its derivatives) have been tested in a number of clinical trials, and this review is focused on the use of MN-based clinical trials. The papers reporting on MN-based clinical trials were retrieved in MedLine up to July 2014, and evaluated for the following endpoints: (a) treated diseases; (b) dosages, number of enrolled patients and duration of treatment; (c) trial success for each MN or MN combinations as reported by authors. The reports satisfying the above endpoints included total numbers of trials and frequencies of randomized, controlled studies, i.e., 81 trials testing ALA, 107 reports testing CoQ10, and 74 reports testing CARN, while only 7 reports were retrieved testing double MN associations, while no report was found testing a triple MN combination. A total of 28 reports tested MN associations with “classical” antioxidants, such as antioxidant nutrients or drugs. Combinations of MN showed better outcomes than individual MN, suggesting forthcoming clinical studies. The criteria in study design and monitoring MN-based clinical trials are discussed

Pharmacological Chaperones and Coenzyme Q10 Treatment Improves Mutant β-Glucocerebrosidase Activity and Mitochondrial Function in Neuronopathic Forms of Gaucher Disease

Gaucher disease (GD) is caused by mutations in the GBA1 gene, which encodes lysosomal β-glucocerebrosidase. Homozygosity for the L444P mutation in GBA1 is associated with high risk of neurological manifestations which are not improved by enzyme replacement therapy. Alternatively, pharmacological chaperones (PCs) capable of restoring the correct folding and trafficking of the mutant enzyme represent promising alternative therapies.Here, we report on how the L444P mutation affects mitochondrial function in primary fibroblast derived from GD patients. Mitochondrial dysfunction was associated with reduced mitochondrial membrane potential, increased reactive oxygen species (ROS), mitophagy activation and impaired autophagic flux.Both abnormalities, mitochondrial dysfunction and deficient β-glucocerebrosidase activity, were partially restored by supplementation with coenzyme Q10 (CoQ) or a L-idonojirimycin derivative, N-[N’-(4-adamantan-1-ylcarboxamidobutyl)thiocarbamoyl]-1,6-anhydro-L-idonojirimycin (NAdBT-AIJ), and more markedly by the combination of both treatments. These data suggest that targeting both mitochondria function by CoQ and protein misfolding by PCs can be promising therapies in neurological forms of GD.España, Ministerio de Sanidad FIS PI13/00129España, Ministerio de Economía y Competitividad SAF2013-44021-R and CTQ2010-15848España, Junta de Andalucía CTS-5725 and FQM-146

Utility of Periodontal exploration in patients with Fibromyalgia

Objetive: Fibromyalgia (FM) is a chronic pain syndrome with unknown etiology, which affects predominantly women. Mitochondrial alteration could have a role in the pathophysilogical mechanisms of inflammatory conditions as FM and periodontitis. The aim of the present study was assay the relationship between both diseases and mitochondrial dysfunction. Patients and methods: We study the presence of periodontitis in twelve patients diagnosed of FM and mitochondrial dysfunction described. The diagnosis of FM was established according to ACR criteria and clinical symptoms were evaluated using the Fibromyalgia Impact Questionnaire (FIQ) and Beck Depression Inventory (BDI). Results: Only one patients of twelve included and agreed to participate in the study were diagnosed with periodontitis. Conclusions: Pending studies with larger numbers of patients, we can conclude that mitochondrial dysfunction in FM is a itself event not related with periodontitis. Periodontitis could be considered a exclusion criterion in all studies about mitochondrial dysfunction in patient

Inhibition of the NLRP3 inflammasome improves lifespan in animal murine model of Hutchinson–Gilford Progeria

Inflammation is a hallmark of aging and accelerated aging syndromes such as Hutchinson–Gilford progeria syndrome (HGPS). In this study, we present evidence of increased expression of the components of the NLRP3 inflammasome pathway in HGPS skin fibroblasts, an outcome that was associated with morphological changes of the nuclei of the cells. Lymphoblasts from HGPS patients also showed increased basal levels of NLRP3 and caspase 1. Consistent with these results, the expression of caspase 1 and Nlrp3, but not of the other inflammasome receptors was higher in the heart and liver of Zmpste24−/− mice, which phenocopy the human disease. These data were further corroborated in LmnaG609G/G609G mice, another HGPS animal model. We also showed that pharmacological inhibition of the NLRP3 inflammasome by its selective inhibitor, MCC950, improved cellular phenotype, significantly extended the lifespan of progeroid animals, and reduced inflammasome-dependent inflammation. These findings suggest that inhibition of the NLRP3 inflammasome is a potential therapeutic approach for the treatment of HGPS.Junta de Andalucía PI-0036-201

Acute oxidant damage promoted on cancer cells by amitriptyline in comparison with some common chemotherapeutic drugs

Oxidative therapy is a relatively new anticancer strategy based on the induction of high levels of oxidative stress, achieved by increasing intracellular reactive oxygen species (ROS) and/or by depleting the protective antioxidant machinery of tumor cells. We focused our investigations on the antitumoral potential of amitriptyline in three human tumor cell lines: H460 (lung cancer), HeLa (cervical cancer), and HepG2 (hepatoma); comparing the cytotoxic effect of amitriptyline with three commonly used chemotherapeutic drugs: camptothecin, doxorubicin, and methotrexate. We evaluated apoptosis, ROS production, mitochondrial mass and activity, and antioxidant defenses of tumor cells. Our results show that amitriptyline produces the highest cellular damage, inducing high levels of ROS followed by irreversible serious mitochondrial damage. Interestingly, an unexpected decrease in antioxidant machinery was observed only for amitriptyline. In conclusion, based on the capacity of generating ROS and inhibiting antioxidants in tumor cells, amitriptyline emerges as a promising new drug to be tested for anticancer therapy