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Mitochondrial dysfunction and mitophagy activation in blood mononuclear cells of fibromyalgia patients: implications in the pathogenesis of the disease.

Cordero MD, De Miguel M, Moreno Fernández AM, Carmona López IM, Garrido Maraver J, Cotán D, Gómez Izquierdo L, Bonal P, Campa F, Bullon P, Navas P, Sánchez Alcázar JA - Arthritis Res. Ther. (2010)

Bottom Line: Mitophagy was confirmed by measuring citrate synthase activity and electron microscopy examination of blood mononuclear cells.We found reduced levels of coenzyme Q10, decreased mitochondrial membrane potential, increased levels of mitochondrial superoxide in blood mononuclear cells, and increased levels of lipid peroxidation in both blood mononuclear cells and plasma from fibromyalgia patients.Mitochondrial dysfunction was also associated with increased expression of autophagic genes and the elimination of dysfunctional mitochondria with mitophagy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centro Andaluz de Biología del Desarrollo (CABD), Universidad Pablo de Olavide-CSIC, Ctra, de Utrera, km, 1, ISCIII, Sevilla 41013, Spain. mdcormor@upo.es

ABSTRACT

Introduction: Fibromyalgia is a chronic pain syndrome with unknown etiology. Recent studies have shown some evidence demonstrating that oxidative stress may have a role in the pathophysiology of fibromyalgia. However, it is still not clear whether oxidative stress is the cause or the effect of the abnormalities documented in fibromyalgia. Furthermore, the role of mitochondria in the redox imbalance reported in fibromyalgia also is controversial. We undertook this study to investigate the role of mitochondrial dysfunction, oxidative stress, and mitophagy in fibromyalgia.

Methods: We studied 20 patients (2 male, 18 female patients) from the database of the Sevillian Fibromyalgia Association and 10 healthy controls. We evaluated mitochondrial function in blood mononuclear cells from fibromyalgia patients measuring, coenzyme Q10 levels with high-performance liquid chromatography (HPLC), and mitochondrial membrane potential with flow cytometry. Oxidative stress was determined by measuring mitochondrial superoxide production with MitoSOX and lipid peroxidation in blood mononuclear cells and plasma from fibromyalgia patients. Autophagy activation was evaluated by quantifying the fluorescence intensity of LysoTracker Red staining of blood mononuclear cells. Mitophagy was confirmed by measuring citrate synthase activity and electron microscopy examination of blood mononuclear cells.

Results: We found reduced levels of coenzyme Q10, decreased mitochondrial membrane potential, increased levels of mitochondrial superoxide in blood mononuclear cells, and increased levels of lipid peroxidation in both blood mononuclear cells and plasma from fibromyalgia patients. Mitochondrial dysfunction was also associated with increased expression of autophagic genes and the elimination of dysfunctional mitochondria with mitophagy.

Conclusions: These findings may support the role of oxidative stress and mitophagy in the pathophysiology of fibromyalgia.

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Related in: MedlinePlus

Autophagic markers in blood mononuclear cells (BMCs) from fibromyalgia (FM) patients. (a) Quantification of acidic vacuoles in control and patient BMCs by LysoTracker fluorescence and flow-cytometry analysis. (b) Reduction of LysoTracker fluorescence in BMCs from FM patients under CoQ10 supplementation (100 μmol/L) for 24 h. Data represent the mean ± SD of three separate experiments. *P < 0.001 between controls and FM patients.
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Figure 4: Autophagic markers in blood mononuclear cells (BMCs) from fibromyalgia (FM) patients. (a) Quantification of acidic vacuoles in control and patient BMCs by LysoTracker fluorescence and flow-cytometry analysis. (b) Reduction of LysoTracker fluorescence in BMCs from FM patients under CoQ10 supplementation (100 μmol/L) for 24 h. Data represent the mean ± SD of three separate experiments. *P < 0.001 between controls and FM patients.

Mentions: Recently, it was demonstrated that CoQ10-deficient fibroblasts exhibit increased levels of lysosomal markers (β-galactosidase, cathepsin, LC3, and Lyso Tracker) and enhanced expression of autophagic genes at both transcriptional and translational levels, indicating the presence of autophagy [9]. To verify that CoQ10 deficiency also induces activation of autophagy in BMCs from FM patients, we first quantified levels of acidic vacuoles in BMCs by using Lysotracker fluorescence and flow-cytometry analysis. Acidic vacuoles were significantly increased in patient BMCs with respect to controls (Figure 4a). To elucidate whether autophagy in CoQ10-deficient BMCs could be mitigated by restoring mitochondrial functionality by CoQ10 supplementation, we cultured both control and patient BMCs in the presence of CoQ10 (100 μmol/L) for 24 hours and analyzed them by Lysotracker fluorescence. As is shown in Figure 4b, CoQ10 supplementation drastically reduced the intensity of Lysotracker fluorescence, indicating a reduction in lysosomal activity after CoQ10 treatment.


Mitochondrial dysfunction and mitophagy activation in blood mononuclear cells of fibromyalgia patients: implications in the pathogenesis of the disease.

Cordero MD, De Miguel M, Moreno Fernández AM, Carmona López IM, Garrido Maraver J, Cotán D, Gómez Izquierdo L, Bonal P, Campa F, Bullon P, Navas P, Sánchez Alcázar JA - Arthritis Res. Ther. (2010)

Autophagic markers in blood mononuclear cells (BMCs) from fibromyalgia (FM) patients. (a) Quantification of acidic vacuoles in control and patient BMCs by LysoTracker fluorescence and flow-cytometry analysis. (b) Reduction of LysoTracker fluorescence in BMCs from FM patients under CoQ10 supplementation (100 μmol/L) for 24 h. Data represent the mean ± SD of three separate experiments. *P < 0.001 between controls and FM patients.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2875645&req=5

Figure 4: Autophagic markers in blood mononuclear cells (BMCs) from fibromyalgia (FM) patients. (a) Quantification of acidic vacuoles in control and patient BMCs by LysoTracker fluorescence and flow-cytometry analysis. (b) Reduction of LysoTracker fluorescence in BMCs from FM patients under CoQ10 supplementation (100 μmol/L) for 24 h. Data represent the mean ± SD of three separate experiments. *P < 0.001 between controls and FM patients.
Mentions: Recently, it was demonstrated that CoQ10-deficient fibroblasts exhibit increased levels of lysosomal markers (β-galactosidase, cathepsin, LC3, and Lyso Tracker) and enhanced expression of autophagic genes at both transcriptional and translational levels, indicating the presence of autophagy [9]. To verify that CoQ10 deficiency also induces activation of autophagy in BMCs from FM patients, we first quantified levels of acidic vacuoles in BMCs by using Lysotracker fluorescence and flow-cytometry analysis. Acidic vacuoles were significantly increased in patient BMCs with respect to controls (Figure 4a). To elucidate whether autophagy in CoQ10-deficient BMCs could be mitigated by restoring mitochondrial functionality by CoQ10 supplementation, we cultured both control and patient BMCs in the presence of CoQ10 (100 μmol/L) for 24 hours and analyzed them by Lysotracker fluorescence. As is shown in Figure 4b, CoQ10 supplementation drastically reduced the intensity of Lysotracker fluorescence, indicating a reduction in lysosomal activity after CoQ10 treatment.

Bottom Line: Mitophagy was confirmed by measuring citrate synthase activity and electron microscopy examination of blood mononuclear cells.We found reduced levels of coenzyme Q10, decreased mitochondrial membrane potential, increased levels of mitochondrial superoxide in blood mononuclear cells, and increased levels of lipid peroxidation in both blood mononuclear cells and plasma from fibromyalgia patients.Mitochondrial dysfunction was also associated with increased expression of autophagic genes and the elimination of dysfunctional mitochondria with mitophagy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centro Andaluz de Biología del Desarrollo (CABD), Universidad Pablo de Olavide-CSIC, Ctra, de Utrera, km, 1, ISCIII, Sevilla 41013, Spain. mdcormor@upo.es

ABSTRACT

Introduction: Fibromyalgia is a chronic pain syndrome with unknown etiology. Recent studies have shown some evidence demonstrating that oxidative stress may have a role in the pathophysiology of fibromyalgia. However, it is still not clear whether oxidative stress is the cause or the effect of the abnormalities documented in fibromyalgia. Furthermore, the role of mitochondria in the redox imbalance reported in fibromyalgia also is controversial. We undertook this study to investigate the role of mitochondrial dysfunction, oxidative stress, and mitophagy in fibromyalgia.

Methods: We studied 20 patients (2 male, 18 female patients) from the database of the Sevillian Fibromyalgia Association and 10 healthy controls. We evaluated mitochondrial function in blood mononuclear cells from fibromyalgia patients measuring, coenzyme Q10 levels with high-performance liquid chromatography (HPLC), and mitochondrial membrane potential with flow cytometry. Oxidative stress was determined by measuring mitochondrial superoxide production with MitoSOX and lipid peroxidation in blood mononuclear cells and plasma from fibromyalgia patients. Autophagy activation was evaluated by quantifying the fluorescence intensity of LysoTracker Red staining of blood mononuclear cells. Mitophagy was confirmed by measuring citrate synthase activity and electron microscopy examination of blood mononuclear cells.

Results: We found reduced levels of coenzyme Q10, decreased mitochondrial membrane potential, increased levels of mitochondrial superoxide in blood mononuclear cells, and increased levels of lipid peroxidation in both blood mononuclear cells and plasma from fibromyalgia patients. Mitochondrial dysfunction was also associated with increased expression of autophagic genes and the elimination of dysfunctional mitochondria with mitophagy.

Conclusions: These findings may support the role of oxidative stress and mitophagy in the pathophysiology of fibromyalgia.

Show MeSH
Related in: MedlinePlus