Cytoprotective and Antioxidants in Peroxisomal Neurodegenerative Diseases

Several of the peroxisomal neurodegenerative disorders are the consequence of a specific deficiency of an enzyme or a transporter involved in peroxisomal beta-oxidation of very long chain fatty acids [1,2]. One of the hallmarks in these peroxisomal rare neurodegenerative diseases and in other common demyelinating disorders is the accompanying oxidative damage and neuroinflammation [3]. Compelling data indicates that oxidative stress can activate microglia leading to the overproduction of pro-inflammatory molecules [4,5]. Thus, targeting oxidative stress to limit neuroinflammation may open a new pharmacological therapy window for these still incurable devastating peroxisomal diseases. Here, we present different natural (resveratrol) [6] and synthetic (organoselenides) [7] antioxidant compounds for their capacity of scavenging oxidative stress and in the perspective therapeutic use against oxidative damage in peroxisomal disorders

Effets d’extraits de raquette du cactus -Opuntia ficus indica- sur la modulation du stress oxydant et du processus inflammatoire liés à la déficience en acyl-CoA oxydase 1 et caractérisation d’une lignée cellulaire BV-2 inactivée pour l'ACOX 1

Acyl-CoA oxidase 1 (ACOX1) deficiency is a rare and severe peroxisomal leukodystrophy associated with a very long-chain fatty acids β-oxidation defect. As peroxisomal defects in microglia appear to be a key component of physiopathogenesis in ACOX1 deficiency, we used the murine BV-2 microglial cell line as model: (i) to evaluate the antioxidant and anti-inflammatory properties of Opuntia ficus-indica cactus cladode extracts; (ii) to characterize a new BV-2 ACOX1 deficient cell line recently generated in our laboratory by CRISPR/Cas9 gene editing. In the first part of this work, the microglia activation, achieved by BV-2 cells exposition to four structurally and biologically well-defined lipopolysaccharides (LPS) serotypes, exhibited a LPS structure-related differential effect on fatty acid β-oxidation and antioxidant enzymes in peroxisomes: LPS derived from Escherichia coli diminished ACOX1 activity while LPS from Salmonella minnesota decreased catalase activity. Interestingly, microglial catalase activity activation was obtained by different cactus extracts. This antioxidant effect was accompanied by an anti-inflammatory effect revealed by the reduction of nitric oxide (NO) LPS-dependent production. Our results suggest that cactus extracts may have a neuroprotective activity in activated microglial cells through the induction of peroxisomal antioxidant activity and the inhibition of NO production. In the second part, the characterization of the BV-2 ACOX1 deficient cell line, with allelic mutations, confirmed the absence of ACOX1 protein and enzymatic activity. Although the mutant cells grew more slowly than control cells, they didn’t show any discernible morphological changes. However, catalase activity, due to the peroxisomal H2O2-degrading enzyme, was significantly enhanced. Subcellular fractionation and ultracentrifugation on Nycodenz gradient studies revealed a relative modification in size and density of peroxisomes in ACOX1-deficient cells. Besides this, ACOX1 deficient cells show a profound modification of inflammatory gene expression (IL-1b, IL-4, TNF-) and particularly of CCL2/MCP-1 protein, which is involved in neuroinflammation. This new Acox1-deficient cell line presents the same biochemical changes shown in the human ACOX1 deficiency and represents a promising model to decipher the consequences of a specific microglial peroxisomal β-oxidation defect on peroxisomal functions, oxidative stress, inflammation and cellular functions.La déficience en Acyl-CoA oxydase 1 (ACOX1) est une leucodystrophie peroxysomale rare et sévère associée à un déficit dans la beta-oxydation des acides gras à très long chaine. A cause du rôle clé de ce déficit peroxysomal microglial dans la physiopathogenèse de la déficience en ACOX1, nous avons utilisé la lignée microgliale murine BV-2 comme modèle : (i) pour évaluer les propriétés antioxydantes et anti-inflammatoires des extraits de raquettes du cactus Opuntia ficus-indica ; (ii) pour caractériser une nouvelle lignée BV-2 déficiente en ACOX1 générée récemment dans notre laboratoire par édition génique grâce à la méthode CRISPR-Cas9. Dans la première partie des travaux, les cellules BV-2, activées par exposition à quatre sérotypes de lipopolysaccharides (LPS), montre un lien entre la structure du LPS et l’effet sur la -oxydation des acides gras et les enzymes antioxydantes dans le peroxysome : les LPS dérivant d’Escherichia coli diminuent l’activité ACOX1 alors que les LPS de Salmonella minnesota réduisent l’activité catalase. Remarquablement, les différents extraits de cactus stimulent l’activité catalase. Cet effet antioxydant est accompagné par un effet anti-inflammatoire attesté par la réduction de la production LPS-dépendante d’oxyde nitrique dans les BV-2. Nos résultats suggèrent que les extraits de cactus auraient une activité neuroprotectrice sur les cellules microgliales activées à travers l’induction d’activités anti-oxydantes peroxysomales et l’inhibition de la production de NO. Dans la deuxième partie des travaux, la caractérisation de la lignée BV-2 déficiente en ACOX1, portant des mutations alléliques, confirme l’absence de la protéine et de l’activité ACOX1. Bien que ces cellules aient une croissance plus faible, elles ne montrent pas de modifications morphologiques détectables. Par contre, l’activité catalase, due à l’enzyme peroxysomale dégradant l’H2O2, est augmentée. Les études par fractionnement subcellulaire et par ultracentrifugation en gradient Nycodenz révèlent une modification de la densité et de la taille de peroxysomes. De plus, ces cellules microgliales déficientes montrent une profonde modification de l’expression des gènes liés à l’inflammation (IL-1b, IL-4, TNF-alpha) et particulièrement l’expression de la protéine CCL2/MCP-1 impliquée dans la neuro-inflammation. Cette nouvelle lignée microgliale déficiente en ACOX1 révèlent les mêmes dérégulations biochimiques que celles décrites chez les patients déficients en ACOX1 et représente donc un modèle pour l’étude des conséquences du déficit de la -oxydation peroxysomale dans la microglie sur les fonctions peroxysomales, le stress oxydatif, l’inflammation et les fonctions cellulaires

Effects of extracts from cactus cladode -Opuntia ficus indica- on the modulation of oxidative stress and inflammatory process related to acyl-CoA oxidase 1 deficiency and characterization of a BV-2 cell line inactivated for ACOX 1

La déficience en Acyl-CoA oxydase 1 (ACOX1) est une leucodystrophie peroxysomale rare et sévère associée à un déficit dans la beta-oxydation des acides gras à très long chaine. A cause du rôle clé de ce déficit peroxysomal microglial dans la physiopathogenèse de la déficience en ACOX1, nous avons utilisé la lignée microgliale murine BV-2 comme modèle : (i) pour évaluer les propriétés antioxydantes et anti-inflammatoires des extraits de raquettes du cactus Opuntia ficus-indica ; (ii) pour caractériser une nouvelle lignée BV-2 déficiente en ACOX1 générée récemment dans notre laboratoire par édition génique grâce à la méthode CRISPR-Cas9. Dans la première partie des travaux, les cellules BV-2, activées par exposition à quatre sérotypes de lipopolysaccharides (LPS), montre un lien entre la structure du LPS et l’effet sur la -oxydation des acides gras et les enzymes antioxydantes dans le peroxysome : les LPS dérivant d’Escherichia coli diminuent l’activité ACOX1 alors que les LPS de Salmonella minnesota réduisent l’activité catalase. Remarquablement, les différents extraits de cactus stimulent l’activité catalase. Cet effet antioxydant est accompagné par un effet anti-inflammatoire attesté par la réduction de la production LPS-dépendante d’oxyde nitrique dans les BV-2. Nos résultats suggèrent que les extraits de cactus auraient une activité neuroprotectrice sur les cellules microgliales activées à travers l’induction d’activités anti-oxydantes peroxysomales et l’inhibition de la production de NO. Dans la deuxième partie des travaux, la caractérisation de la lignée BV-2 déficiente en ACOX1, portant des mutations alléliques, confirme l’absence de la protéine et de l’activité ACOX1. Bien que ces cellules aient une croissance plus faible, elles ne montrent pas de modifications morphologiques détectables. Par contre, l’activité catalase, due à l’enzyme peroxysomale dégradant l’H2O2, est augmentée. Les études par fractionnement subcellulaire et par ultracentrifugation en gradient Nycodenz révèlent une modification de la densité et de la taille de peroxysomes. De plus, ces cellules microgliales déficientes montrent une profonde modification de l’expression des gènes liés à l’inflammation (IL-1b, IL-4, TNF-alpha) et particulièrement l’expression de la protéine CCL2/MCP-1 impliquée dans la neuro-inflammation. Cette nouvelle lignée microgliale déficiente en ACOX1 révèlent les mêmes dérégulations biochimiques que celles décrites chez les patients déficients en ACOX1 et représente donc un modèle pour l’étude des conséquences du déficit de la -oxydation peroxysomale dans la microglie sur les fonctions peroxysomales, le stress oxydatif, l’inflammation et les fonctions cellulaires.Acyl-CoA oxidase 1 (ACOX1) deficiency is a rare and severe peroxisomal leukodystrophy associated with a very long-chain fatty acids β-oxidation defect. As peroxisomal defects in microglia appear to be a key component of physiopathogenesis in ACOX1 deficiency, we used the murine BV-2 microglial cell line as model: (i) to evaluate the antioxidant and anti-inflammatory properties of Opuntia ficus-indica cactus cladode extracts; (ii) to characterize a new BV-2 ACOX1 deficient cell line recently generated in our laboratory by CRISPR/Cas9 gene editing. In the first part of this work, the microglia activation, achieved by BV-2 cells exposition to four structurally and biologically well-defined lipopolysaccharides (LPS) serotypes, exhibited a LPS structure-related differential effect on fatty acid β-oxidation and antioxidant enzymes in peroxisomes: LPS derived from Escherichia coli diminished ACOX1 activity while LPS from Salmonella minnesota decreased catalase activity. Interestingly, microglial catalase activity activation was obtained by different cactus extracts. This antioxidant effect was accompanied by an anti-inflammatory effect revealed by the reduction of nitric oxide (NO) LPS-dependent production. Our results suggest that cactus extracts may have a neuroprotective activity in activated microglial cells through the induction of peroxisomal antioxidant activity and the inhibition of NO production. In the second part, the characterization of the BV-2 ACOX1 deficient cell line, with allelic mutations, confirmed the absence of ACOX1 protein and enzymatic activity. Although the mutant cells grew more slowly than control cells, they didn’t show any discernible morphological changes. However, catalase activity, due to the peroxisomal H2O2-degrading enzyme, was significantly enhanced. Subcellular fractionation and ultracentrifugation on Nycodenz gradient studies revealed a relative modification in size and density of peroxisomes in ACOX1-deficient cells. Besides this, ACOX1 deficient cells show a profound modification of inflammatory gene expression (IL-1b, IL-4, TNF-) and particularly of CCL2/MCP-1 protein, which is involved in neuroinflammation. This new Acox1-deficient cell line presents the same biochemical changes shown in the human ACOX1 deficiency and represents a promising model to decipher the consequences of a specific microglial peroxisomal β-oxidation defect on peroxisomal functions, oxidative stress, inflammation and cellular functions

Ultra-Wideband Compact Fractal Antenna for WiMAX, WLAN, C and X Band Applications

In this paper, a compact dual-wideband fractal antenna is created for Bluetooth, WiMAX, WLAN, C, and X band applications. The proposed antenna consists of a circularly shaped resonator that contains square slots and a ground plane where a gap line is incorporated to increase the gain and bandwidth with a small volume of 40 × 34 × 1.6 mm3. The patch was supported by the FR4 dielectric, which had a permittivity of 4.4 and tan δ = 0.02. A 50 Ω microstrip line fed this antenna. The antenna was designed by the HFSS program, and after that, the simulated results were validated using the measured results. The measurement results confirm that the suggested antenna achieves dual-band frequencies ranging from 2.30 to 4.10 GHz, and from 6.10 GHz to 10.0 GHz, resonating at 2.8, 3.51, 6.53, and 9.37 GHz, respectively, for various applications including commercial, scholarly, and medical applications. Moreover, the antenna’s ability to operate within the frequency range of 3.1–10.6 GHz is in accordance with the FCC guidelines for the use of UWB antennas in breast cancer detection. Over the operational bands, the gain varied between 2 and 9 dB, and an efficiency of 92% was attained. A good agreement between the simulation and the measured results was found

Protective Effect of Cactus Cladode Extracts on Peroxisomal Functions in Microglial BV-2 Cells Activated by Different Lipopolysaccharides

In this study, we aimed to evaluate the antioxidant and anti-inflammatory properties of Opuntia ficus-indica cactus cladode extracts in microglia BV-2 cells. Inflammation associated with microglia activation in neuronal injury can be achieved by LPS exposure. Using four different structurally and biologically well-characterized LPS serotypes, we revealed a structure-related differential effect of LPS on fatty acid β-oxidation and antioxidant enzymes in peroxisomes: Escherichia coli-LPS decreased ACOX1 activity while Salmonella minnesota-LPS reduced only catalase activity. Different cactus cladode extracts showed an antioxidant effect through microglial catalase activity activation and an anti-inflammatory effect by reducing nitric oxide (NO) LPS-dependent production. These results suggest that cactus extracts may possess a neuroprotective activity through the induction of peroxisomal antioxidant activity and the inhibition of NO production by activated microglial cells

Peroxisomal acyl-coa oxidase type 1: anti-inflammatory and anti-aging properties with a special emphasis on studies with lps and argan oil as a model transposable to aging

International audienceTo clarify appropriateness of current claims for health and wellness virtues of argan oil, studies were conducted in inflammatory states. LPS induces inflammation with reduction of PGC1-α signaling and energy metabolism. Argan oil protected the liver against LPS toxicity and interestingly enough preservation of peroxisomal acyl-CoA oxidase type 1 (ACOX1) activity against depression by LPS. This model of LPS-driven toxicity circumvented by argan oil along with a key anti-inflammatory role attributed to ACOX1 has been here transposed to model aging. This view is consistent with known physiological role of ACOX1 in yielding precursors of specialized proresolving mediators (SPM) and with characteristics of aging and related disorders including reduced PGC1-α function and improvement by strategies rising ACOX1 (via hormonal gut FGF19 and nordihydroguaiaretic acid in metabolic syndrome and diabetes conditions) and SPM (neurodegenerative disorders, atherosclerosis, and stroke). Delay of aging to resolve inflammation results from altered production of SPM, SPM improving most aging disorders. The strategic metabolic place of ACOX1, upstream of SPM biosynthesis, along with ability of ACOX1 preservation/induction and SPM to improve aging-related disorders and known association of aging with drop in ACOX1 and SPM, all converge to conclude that ACOX1 represents a previously unsuspected and currently emerging antiaging protein

Peroxisomal Acyl-CoA Oxidase Type 1: Anti-Inflammatory and Anti-Aging Properties with a Special Emphasis on Studies with LPS and Argan Oil as a Model Transposable to Aging

To clarify appropriateness of current claims for health and wellness virtues of argan oil, studies were conducted in inflammatory states. LPS induces inflammation with reduction of PGC1-α signaling and energy metabolism. Argan oil protected the liver against LPS toxicity and interestingly enough preservation of peroxisomal acyl-CoA oxidase type 1 (ACOX1) activity against depression by LPS. This model of LPS-driven toxicity circumvented by argan oil along with a key anti-inflammatory role attributed to ACOX1 has been here transposed to model aging. This view is consistent with known physiological role of ACOX1 in yielding precursors of specialized proresolving mediators (SPM) and with characteristics of aging and related disorders including reduced PGC1-α function and improvement by strategies rising ACOX1 (via hormonal gut FGF19 and nordihydroguaiaretic acid in metabolic syndrome and diabetes conditions) and SPM (neurodegenerative disorders, atherosclerosis, and stroke). Delay of aging to resolve inflammation results from altered production of SPM, SPM improving most aging disorders. The strategic metabolic place of ACOX1, upstream of SPM biosynthesis, along with ability of ACOX1 preservation/induction and SPM to improve aging-related disorders and known association of aging with drop in ACOX1 and SPM, all converge to conclude that ACOX1 represents a previously unsuspected and currently emerging antiaging protein

Cytoprotective and Antioxidants in Peroxisomal Neurodegenerative Diseases

Several of the peroxisomal neurodegenerative disorders are the consequence of a specific deficiency of an enzyme or a transporter involved in peroxisomal beta-oxidation of very long chain fatty acids [1,2]. One of the hallmarks in these peroxisomal rare neurodegenerative diseases and in other common demyelinating disorders is the accompanying oxidative damage and neuroinflammation [3]. Compelling data indicates that oxidative stress can activate microglia leading to the overproduction of pro-inflammatory molecules [4,5]. Thus, targeting oxidative stress to limit neuroinflammation may open a new pharmacological therapy window for these still incurable devastating peroxisomal diseases. Here, we present different natural (resveratrol) [6] and synthetic (organoselenides) [7] antioxidant compounds for their capacity of scavenging oxidative stress and in the perspective therapeutic use against oxidative damage in peroxisomal disorders

Protective Effect of Nopal Cactus (<i>Opuntia ficus-indica</i>) Seed Oil against Short-Term Lipopolysaccharides-Induced Inflammation and Peroxisomal Functions Dysregulation in Mouse Brain and Liver

Exposure to endotoxins (lipopolysaccharides, LPS) may lead to a potent inflammatory cytokine response and a severe impairment of metabolism, causing tissue injury. The protective effect provided by cactus seed oil (CSO), from Opuntia ficus-indica, was evaluated against LPS-induced inflammation, dysregulation of peroxisomal antioxidant, and β-oxidation activities in the brain and the liver. In both tissues, a short-term LPS exposure increased the proinflammatory interleukine-1β (Il-1β), inducible Nitroxide synthase (iNos), and Interleukine-6 (Il-6). In the brain, CSO action reduced only LPS-induced iNos expression, while in the liver, CSO attenuated mainly the hepatic Il-1β and Il-6. Regarding the peroxisomal antioxidative functions, CSO treatment (as Olive oil (OO) or Colza oil (CO) treatment) induced the hepatic peroxisomal Cat gene. Paradoxically, we showed that CSO, as well as OO or CO, treatment can timely induce catalase activity or prevent its induction by LPS, respectively, in both brain and liver tissues. On the other hand, CSO (as CO) pretreatment prevented the LPS-associated Acox1 gene and activity decreases in the liver. Collectively, CSO showed efficient neuroprotective and hepato-protective effects against LPS, by maintaining the brain peroxisomal antioxidant enzyme activities of catalase and glutathione peroxidase, and by restoring hepatic peroxisomal antioxidant and β-oxidative capacities