ABSTRACT
Hypermethioninemia is a disorder characterized by high plasma levels of methionine (Met) and its metabolites such as methionine sulfoxide (MetO). Studies have reported associated inflammatory complications, but the mechanisms involved in the pathophysiology of hypermethioninemia are still uncertain. The present study aims to evaluate the effect of chronic administration of Met and/or MetO on phenotypic characteristics of macrophages, in addition to oxidative stress, purinergic system, and inflammatory mediators in macrophages. In this study, Swiss male mice were subcutaneously injected with Met and MetO at concentrations of 0.35-1.2 g/kg body weight and 0.09-0.3 g/kg body weight, respectively, from the 10th-38th day post-birth, while the control group was treated with saline solution. The results revealed that Met and/or MetO induce an M1/classical activation phenotype associated with increased levels of tumor necrosis factor alpha and nitrite, and reduced arginase activity. It was also found that Met and/or MetO alter the activity of antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase, as well as the levels of thiol and reactive oxygen species in macrophages. The chronic administration of Met and/or MetO also promotes alteration in the hydrolysis of ATP and ADP, as indicated by the increased activity of ectonucleotidases. These results demonstrate that chronic administration of Met and/or MetO promotes activated pro-inflammatory profile by inducing M1/classical macrophage polarization. Thus, the changes in redox status and purinergic system upon chronic Met and/or MetO exposure may contribute towards better understanding of the alterations consistent with hypermethioninemic patients.
Subject(s)
Amino Acid Metabolism, Inborn Errors/immunology , Glycine N-Methyltransferase/deficiency , Macrophages/immunology , Methionine/analogs & derivatives , Animals , Catalase/metabolism , Cell Polarity , Glutathione Peroxidase/metabolism , Glycine N-Methyltransferase/immunology , Macrophages/drug effects , Male , Methionine/administration & dosage , Methionine/metabolism , Methionine/pharmacology , Mice , Oxidation-Reduction , Oxidative Stress , Phenotype , Superoxide Dismutase/metabolismABSTRACT
Glioblastoma is a devastating tumor affecting the central nervous system with infiltrative capacity, high proliferation rate and chemoresistance. Therefore, it is urgent to find new therapeutic alternatives that improve this prognosis. Herein, we focused on tannic acid (TA) a polyphenol with antioxidant and antiproliferative activities. In this work, the antitumor and antioxidant effects of TA on rat (C6) glioblastoma cells and their cytotoxicity relative to primary astrocyte cultures were evaluated in vitro. Cells were exposed to TA of 6.25 to 75 µM for 24, 48 and/or 72 h. In addition, colony formation, migration and cell adhesion were analyzed and flow cytometry was used to analyze cell death and cell cycle. Next, the action of TA was evaluated in a preclinical glioblastoma model performed on Wistar rats. In this protocol, the animals were treated with a dose of 50 mg/kg/day TA for 15 days. Our results demonstrated that TA induced in vitro selective antiglioma activity, not demonstrating cytotoxicity in astrocyte culture. It induced cell death by apoptosis and cell cycle arrest, reducing formation and size of colonies, cell migration/adhesion and showing to be a potential antioxidant. Interestingly, the antiglioma effect was also observed in vivo, as TA decreased tumor volume by 55%, accompanied by an increase in the area of intratumoral necrosis and infiltration of lymphocytes without causing systemic damage. To the best of our knowledge, this is the first study to report TA activity in a GBM preclinical model. Thus, this natural compound is promising as a treatment for glioblastoma.
Subject(s)
Antineoplastic Agents/therapeutic use , Brain Neoplasms/drug therapy , Glioblastoma/drug therapy , Tannins/therapeutic use , Xenograft Model Antitumor Assays/methods , Animals , Antineoplastic Agents/pharmacology , Brain Neoplasms/pathology , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Proliferation/physiology , Cell Survival/drug effects , Cell Survival/physiology , Dose-Response Relationship, Drug , Female , Glioblastoma/pathology , Male , Rats , Rats, Wistar , Tannins/pharmacologyABSTRACT
The present study describes the synthesis of a novel series of thiazolidin-4-one and thiazinan-4-one using 1-(2-aminoethyl)pyrrolidine as amine precursor. All compounds were synthesised by one-pot three component cyclocondensation reaction from the amine, a substituted benzaldehyde and a mercaptocarboxylic acid. The compounds were obtained in moderate to good yields and were identified and characterised by 1H, 13 C, 2 D NMR and GC/MS techniques. The compounds also were screened for their in vitro acetylcholinesterase (AChE) activity in hippocampus and cerebral cortex on Wistar rats. The six most potent compounds have been investigated for their cytotoxicity by cell viability assay of astrocyte primary culture, an important cell of central nervous system. We highlighted two compounds (6a and 6k) that had the lowest IC50 in hippocampus (5.20 and 4.46 µM) and cerebral cortex (7.40 and 6.83 µM). These preliminary and important results could be considered a starting point for the development of new AChE inhibitory agents.
Subject(s)
Acetylcholinesterase/metabolism , Cholinesterase Inhibitors/pharmacology , Thiazines/pharmacology , Thiazolidines/pharmacology , Animals , Cerebral Cortex/drug effects , Cerebral Cortex/enzymology , Cholinesterase Inhibitors/chemical synthesis , Cholinesterase Inhibitors/chemistry , Dose-Response Relationship, Drug , Hippocampus/drug effects , Hippocampus/enzymology , Molecular Structure , Rats , Rats, Wistar , Structure-Activity Relationship , Thiazines/chemical synthesis , Thiazines/chemistry , Thiazolidines/chemical synthesis , Thiazolidines/chemistryABSTRACT
Introduction: Glioblastoma (GB) is the most common malignant brain tumor and is characterized by high invasiveness, poor prognosis, and limited therapeutic options. Silencing gene expression, through the use of small interfering RNA (siRNA), has been proposed as an alternative to conventional cancer therapy. Here, we evaluated the potential of CD73 as a new therapeutic target, since it is overexpressed in solid tumors and has emerged as a promising target to control GB progression.Methods: A cationic nanoemulsion (NE) as an intravenous siRNA-CD73 delivery system was developed and its effect on C6 glioma cell viability was determined.Results: The nanostructured system was effective in complexing oligonucleotides for delivery to target cells. In addition, we observed that the NE-siRNA-CD73 complex was effective in reducing CD73 protein levels and AMPase activity, which were related to decreased C6 glioma cell viability.Conclusions: These findings indicate the potential of siRNA-CD73-loaded cationic NE as a therapeutic alternative for glioma treatment.
Subject(s)
5'-Nucleotidase/genetics , Glioma/therapy , RNA, Small Interfering/administration & dosage , RNA, Small Interfering/therapeutic use , Animals , Astrocytes/cytology , Astrocytes/metabolism , Cations/chemistry , Cell Line, Tumor , Cells, Cultured , Drug Carriers/chemistry , Emulsions/chemistry , Glioma/genetics , RNA, Small Interfering/genetics , RNAi Therapeutics , RatsABSTRACT
A series of nineteen benzothiazin-4-ones from N-(3-aminopropyl) piperidine, 4-(2-aminoethyl)morpholine or 1-(2-aminoethyl)piperidine, aliphatic or aromatic aldehyde and thiosalicylic acid, were synthesized in good yields by multicomponent one-pot reactions. The solvent was toluene and this efficient procedure afforded the desired heterocycles in 5 h. Identification and characterization were achieved by NMR and GC-MS techniques. In vitro AChE activities of all compounds were evaluated in cerebral cortex and hippocampus of rats and in general, the results in cortex were more promising than hippocampus. The benzothiazinone 5Bd showed the best AChE inhibition activity IC50 8.48 µM (cortex) and IC50 39.80 µM (hippocampus). The cytotoxicity of seven compounds in MCR-5 human fibroblast cell by SRB test in 24 h were evaluated and 5Bd suggest preliminary safety, showing no cytotoxicity at 100 µM. Finally, these important findings could be a starting point for the development of new AChE inhibitors agents and will provide the basis for new studies.
Subject(s)
Acetylcholinesterase/metabolism , Benzothiadiazines/pharmacology , Cholinesterase Inhibitors/pharmacology , Animals , Benzothiadiazines/chemical synthesis , Benzothiadiazines/chemistry , Cells, Cultured , Cholinesterase Inhibitors/chemical synthesis , Cholinesterase Inhibitors/chemistry , Dose-Response Relationship, Drug , Drug Design , Fibroblasts/drug effects , Humans , Male , Molecular Structure , Rats , Rats, Wistar , Structure-Activity RelationshipABSTRACT
Methionine is an essential amino acid involved in critical metabolic process, and regulation of methionine flux through metabolism is important to supply this amino acid for cell needs. Elevation in plasma methionine commonly occurs due to mutations in methionine-metabolizing enzymes, such as methionine adenosyltransferase. Hypermethioninemic patients exhibit clinical manifestations, including neuronal and liver disorders involving inflammation and tissue injury, which pathophysiology is not completely established. Here, we hypothesize that alterations in macrophage inflammatory response may contribute to deleterious effects of hypermethioninemia. To this end, macrophage primary cultures were exposed to methionine (1 mM) and/or its metabolite methionine sulfoxide (0.5 mM), and M1/proinflammatory or M2/anti-inflammatory macrophage polarization was evaluated. In addition, inflammation-related pathways including oxidative stress parameters, as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities; reactive oxygen species (ROS) production, and purinergic signaling, as ATP/ADP/AMPase activities, were investigated. Methionine and/or methionine sulfoxide induced M1/classical macrophage activation, which is related to proinflammatory responses characterized by increased iNOS activity and TNF-α release. Further experiments showed that treatments promoted alterations on redox state of macrophages by differentially modulated SOD and CAT activities and ROS levels. Finally, methionine and/or methionine sulfoxide treatment also altered the extracellular nucleotide metabolism, promoting an increase of ATPase/ADPase activities in macrophages. In conclusion, these findings contribute to better understand the participation of proinflammatory responses in cell injury observed in hypermethioninemic patients.
Subject(s)
Macrophages/metabolism , Methionine/analogs & derivatives , Methionine/pharmacology , Oxidative Stress/drug effects , Signal Transduction/drug effects , Animals , Catalase/metabolism , Glutathione Peroxidase/metabolism , Male , Mice , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolismABSTRACT
BACKGROUND: Glioblastomas are the most devastating brain tumor characterized by chemoresistance development and poor prognosis. Macrophages are a component of tumor microenvironment related to glioma malignancy. The relation among inflammation, innate immunity and cancer is accepted; however, molecular and cellular mechanisms mediating this relation and chemoresistance remain unresolved. OBJECTIVE: Here we evaluated whether glioma sensitive or resistant to temozolomide (TMZ) modulate macrophage polarization and inflammatory pathways associated. The impact of glioma-macrophage crosstalk on glioma proliferation was also investigated. METHODS: GL261 glioma chemoresistance was developed by exposing cells to increasing TMZ concentrations over a period of 6months. Mouse peritoneal macrophages were exposed to glioma-conditioned medium or co-cultured directly with glioma sensitive (GL) or chemoresistant (GLTMZ). Macrophage polarization, in vitro and in vivo glioma proliferation, redox parameters, ectonucleotidase activity and ATP cytotoxicity were performed. RESULTS: GLTMZ cells were more effective than GL in induce M2-like macrophage polarization and in promote a strong immunosuppressive environment characterized by high IL-10 release and increased antioxidant potential, which may contribute to glioma chemoresistance and proliferation. Interestingly, macrophage-GLTMZ crosstalk enhanced in vitro and in vivo proliferation of chemoresistant cells, decreased ectonucleotidase activities, which was followed by increased macrophage sensitivity to ATP induced death. CONCLUSIONS: Results suggest a differential macrophage modulation by GLTMZ cells, which may favor the maintenance of immunosuppressive tumor microenvironment and glioma proliferation. GENERAL SIGNIFICANCE: The induction of immunosuppressive environment and macrophage education by chemoresistant gliomas may be important for tumor recovery after chemotherapy and could be considered to overcome chemoresistance development.
Subject(s)
Dacarbazine/analogs & derivatives , Drug Resistance, Neoplasm/genetics , Glioma/drug therapy , Inflammation/drug therapy , Animals , Antineoplastic Agents, Alkylating/administration & dosage , Antioxidants/metabolism , Apoptosis/drug effects , Cell Line, Tumor , Cell Polarity/drug effects , Dacarbazine/administration & dosage , Disease Models, Animal , Glioma/metabolism , Glioma/pathology , Humans , Inflammation/metabolism , Inflammation/pathology , Macrophages/drug effects , Macrophages/metabolism , Mice , Receptors, Purinergic/genetics , Temozolomide , Tumor Microenvironment/drug effectsABSTRACT
Glioblastoma multiforme (GBM) is the worst and most common brain tumor, characterized by high proliferation and invasion rates. Nanoparticles of biodegradable polymers for anticancer drug delivery have attracted interest in recent years since they provide targeted delivery and may overcame the obstacle imposed by blood-brain barrier. Here we investigated the antitumoral effect of ketoprofen-loaded nanocapsules (Keto-NC) treatment on in vitro and in vivo glioma progression. We observed that Keto-NC treatment decreased selectively the cell viability of a panel of glioma cell lines, while did not exhibited toxicity to astrocytes. We further demonstrate that the treatment with sub-therapeutic dose of Keto-NC reduced the in vivo glioma growth as well as reduced the malignity characteristics of implanted tumors. Keto-NC treatment improved the weight, the locomotion/exploration behavior of glioma-bearing rats. Importantly, Keto-NC treatment neither induced mortality or peripheral damage. Finally, Ketoprofen also altered the extracellular nucleotide metabolism of peripheral lymphocytes, suggesting that antiinflammatory effects of ketoprofen could also be associated with the modulation of the adenine nucleotide metabolism in lymphocytes. Data indicate at first time the potential of Keto-NC as a promising therapeutic alterative to GBM treatment.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/administration & dosage , Antineoplastic Agents/administration & dosage , Brain Neoplasms/drug therapy , Glioma/drug therapy , Ketoprofen/administration & dosage , Nanocapsules/administration & dosage , Animals , Brain Neoplasms/pathology , Cell Line, Tumor , Disease Models, Animal , Glioma/pathology , Humans , Male , Rats , Rats, Wistar , Tumor Burden/drug effectsABSTRACT
The present study aimed to investigate the influence of N-acetylcysteine (NAC) on cadmium (Cd) poisoning by evaluating Cd concentration in tissues, hematological indices as well as the activity of NTPDase, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes of rats exposed to Cd and co-treated with NAC. For this purpose, the rats received Cd (2 mg/kg) and NAC (150 mg/kg) by gavage every other day for 30 days. Animals were divided into four groups (n = 6-8): control/saline, NAC, Cd, and Cd/NAC. Cd exposure increased Cd concentration in plasma, spleen and thymus, and NAC co-treatment modulated this augment in both lymphoid organs. Cd exposure reduced red blood cell count, hemoglobin content and hematocrit value. Cd intoxication caused a decrease in total white blood cell count. NAC treatment per se caused an increase in lymphocyte and a decrease in neutrophil counts. On contrary, Cd exposure caused a decrease in lymphocyte and an increase in neutrophil and monocyte counts. NAC reversed or ameliorated the hematological impairments caused by Cd poisoning. There were no significant alterations in the NTPDase activity in lymphocytes of rats treated with Cd and/or NAC. Cd caused a decrease in the activities of lymphocyte AChE, whole blood AChE and serum BChE. However, NAC co-treatment was inefficient in counteracting the negative effect of Cd in the cholinesterase activities. The present investigation provides ex vivo evidence supporting the hypothesis that Cd induces immunotoxicity by interacting with the lymphoid organs, altering hematological parameters and inhibiting peripheral cholinesterase activity. Also, it highlights the possibility to use NAC as adjuvant against toxicological conditions.
Subject(s)
Acetylcholinesterase/metabolism , Acetylcysteine/pharmacology , Antigens, CD/metabolism , Apyrase/metabolism , Butyrylcholinesterase/metabolism , Cadmium/pharmacology , Acetylcholinesterase/blood , Acetylcysteine/administration & dosage , Animals , Antigens, CD/blood , Apyrase/antagonists & inhibitors , Apyrase/blood , Butyrylcholinesterase/blood , Cadmium/administration & dosage , Cadmium/blood , Lymphocytes/drug effects , Lymphocytes/enzymology , Lymphocytes/metabolism , Male , Rats , Rats, Wistar , Structure-Activity RelationshipABSTRACT
The aim of this study is to evaluate the role of cholinesterases as an inflammatory marker in acute and chronic infection by Trypanosoma evansi in rabbits experimentally infected. Twelve adult female New Zealand rabbits were used and divided into two groups with 6 animals each: control group (rabbits 1-6) and infected group (rabbits 7-12). Infected group received intraperitoneally 0.5 mL of blood from a rat containing 108 parasites per animal. Blood samples used for cholinesterases evaluation were collected on days 0, 2, 7, 12, 27, 42, 57, 87, 102 and 118 days post-inoculation (PI). Increased activity (P<0.05) of butyrylcholinesterase (BChE) and acetylcholinesterase (AChE) were observed in the blood on days 7 and 27, respectively and no differences were observed in cholinesterase activity in other periods. No significant difference in AChE activity (P>0.05) was observed in the encephalic structures. The increased activities of AChE and BChE probably have a pro-inflammatory purpose, attempting to reduce the concentration of acetylcholine, a neurotransmitter which has an anti-inflammatory property. Therefore, cholinesterase may be inflammatory markers in infection with T. evansi in rabbits.
Subject(s)
Acetylcholinesterase/blood , Butyrylcholinesterase/blood , Trypanosomiasis/enzymology , Acute Disease , Animals , Biomarkers/blood , Chronic Disease , Female , Parasitemia/blood , Rabbits , RatsABSTRACT
The aim of this study was to investigate the ability of tannic acid (TA) in preventing memory deficits and neurochemical alterations observed in a model for Sporadic Dementia of Alzheimer's Type. Rats were treated with TA (30 mg/kg) daily for 21 days, and subsequently received intracerebroventricular injection of streptozotocin (STZ). We observed that STZ induced learning and memory impairments; however, treatment with TA was able to prevent these effects. In cerebral cortex and hippocampus, STZ induced an increase in acetylcholinesterase activity, reduced Na+, K+-ATPase activity and induced oxidative stress increasing thiobarbituric acid-reactive substances, nitrites and reactive oxygen species levels and reducing the activity of antioxidant enzymes. Treatment with TA was able in prevent the major of these neurochemical alterations. In conclusion, TA prevented memory deficits, alterations in brain enzyme activities, and oxidative damage induced by STZ. Thus, TA can be an interesting strategy in the prevention of Sporadic Alzheimer's Disease.
Subject(s)
Alzheimer Disease , Acetylcholinesterase/metabolism , Adenosine Triphosphatases , Alzheimer Disease/chemically induced , Alzheimer Disease/drug therapy , Alzheimer Disease/prevention & control , Animals , Disease Models, Animal , Maze Learning , Oxidation-Reduction , Oxidative Stress , Rats , Rats, Wistar , Streptozocin/toxicity , TanninsABSTRACT
Al adjuvants are used in vaccines to increase the immune response. NTPDase and AChE play a pivotal role and act in the regulation of the immune system. The effect of Al exposure in vitro and in vivo on NTPDase and AChE activities in the lymphocytes of rats was determined. In vitro, ATP hydrolysis was decreased by 20.4% and 17.3% and ADP hydrolysis was decreased by 36.5% and 34.8%, in groups D and E, respectively, when compared to the control. AChE activity was increased by 157.3%, 152.5%, 74.7% and 90.8% in groups B, C, D, and E, respectively, when compared to the control. In vivo, ATP hydrolysis was increased by 85% and 86% and ADP hydrolysis was increased by 104.2% and 74%, in Al plus citrate and Al groups, respectively, when compared to the control. AChE activity was increased by 50.7% in Al plus citrate and by 28.6% in Al groups, when compared to the control. Our results show that Al exposure both in vitro and in vivo altered NTPDase and AChE activities in lymphocytes. These results may demonstrate the ability of Al to elicit the immune system, where NTPDase and AChE activities can act as purinergic and cholinergic markers in lymphocytes.
Subject(s)
Acetylcholinesterase/metabolism , Aluminum/pharmacology , Antigens, CD/metabolism , Apyrase/metabolism , Lymphocyte Activation/drug effects , T-Lymphocytes/immunology , T-Lymphocytes/metabolism , Adenosine Triphosphatases/metabolism , Animals , Cells, Cultured , Enzyme Activation/drug effects , GPI-Linked Proteins , Male , Protein Binding , RatsABSTRACT
A membrane of cationic starch-derivative/poly(vinyl alcohol) was prepared and utilized as a support to immobilize a ß-cyclodextrin/curcumin inclusion complex. The resulting material (denote as ß-CD/CUR-MBN) was characterized in detail by different techniques. In vitro experiments revealed that ß-CD/CUR-MBN enables the controlling of the curcumin release process, which is guided by the relaxation of the polymer matrix. Moreover, cytotoxic assays were performed to investigate the effect of ß-CD/CUR-MBN on two cancer cell lines (melanoma and glioblastoma). The results showed that the polymeric membrane exerts higher cytotoxicity against these cells than free curcumin. Also, ß-CD/CUR-MBN exerted a prolonged cytotoxic effect (up to 96 h), even using a low concentration (50 µg mL-1), indicating that the curcumin in the polymeric membrane showed increased bioavailability under the tested condition. ß-CD/CUR-MBN was non-cytotoxic against normal cells suggesting a specific action of this material against target cancer cells. The results reported here allow ranks ß-CD/CUR-MBN as a promising biomaterial to act as a local drug delivery system to treat cancer.
Subject(s)
Antineoplastic Agents/pharmacology , Cations/chemistry , Curcumin/pharmacology , Melanoma/drug therapy , Starch/chemistry , beta-Cyclodextrins/chemistry , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacokinetics , Biological Availability , Cell Line, Tumor , Curcumin/chemistry , Curcumin/pharmacokinetics , HumansABSTRACT
Bipolar disorder is a chronic mood disorder characterized by episodes of mania and depression. The aim of this study was to investigate the effects of blackberry extract on behavioral parameters, oxidative stress and inflammatory markers in a ketamine-induced model of mania. Animals were pretreated with extract (200â¯mg/kg, once a day for 14 days), lithium chloride (45â¯mg/kg, twice a day for 14 days), or vehicle. Between the 8th and 14th days, the animals received an injection of ketamine (25â¯mg/kg) or vehicle. On the 15th day, thirty minutes after ketamine administration, the animals' locomotion was assessed using open-field apparatus. After the experiments, the animals were euthanized and cerebral structures were removed for neurochemical analyses. The results showed that ketamine treatment induced hyperlocomotion and oxidative damage in the cerebral cortex, hippocampus and striatum. In contrast, pretreatment with the extract or lithium was able to prevent hyperlocomotion and oxidative damage in the cerebral cortex, hippocampus, and striatum. In addition, IL-6 and IL-10 levels were increased by ketamine, while the extract prevented these effects in the cerebral cortex. Pretreatment with the extract was also effective in decreasing IL-6 and increasing the level of IL-10 in the striatum. In summary, our findings suggest that blackberry consumption could help prevent or reduce manic episodes, since this extract have demonstrated neuroprotective properties as well as antioxidant and anti-inflammatory effects in the ketamine-induced mania model.
Subject(s)
Anthocyanins , Fruit , Mania/metabolism , Plant Extracts/pharmacology , Rubus , Animals , Antimanic Agents/pharmacology , Behavior, Animal/drug effects , Catalase/drug effects , Catalase/metabolism , Cerebral Cortex/drug effects , Cerebral Cortex/metabolism , Cytokines/drug effects , Cytokines/metabolism , Disease Models, Animal , Excitatory Amino Acid Antagonists/toxicity , Glutathione Peroxidase/drug effects , Glutathione Peroxidase/metabolism , Hippocampus/drug effects , Hippocampus/metabolism , Ketamine/toxicity , Lithium Chloride/pharmacology , Mania/chemically induced , Mania/physiopathology , Neostriatum/drug effects , Neostriatum/metabolism , Open Field Test , Plant Extracts/chemistry , Rats , Superoxide Dismutase/drug effects , Superoxide Dismutase/metabolism , Thiobarbituric Acid Reactive Substances/metabolismABSTRACT
Tannic acid (TA) is a hydrolysable glycosidic polyphenol polymer of gallic acid, which possesses neuroprotective properties. The aim of this study was to evaluate the effect of TA treatment on cognitive performance and neurochemical changes in an experimental model of sporadic dementia of Alzheimer's type (SDAT) induced by intracerebroventricular (ICV) injection of streptozotocin (STZ) and to explore the potential cellular and molecular mechanisms underlying these effects. Adult male rats were divided into four groups: control, TA, STZ, and TA + STZ. Animals from TA and TA + STZ groups were treated with TA (30 mg/kg) daily, by gavage, for 21 days; others groups received water (1 mL/kg). Subsequently, an ICV injection of STZ (3 mg/kg) was administered into the lateral ventricles of animals from STZ and TA + STZ groups, while other groups received citrate buffer. Cognitive deficits (short-term memory), neuronal survival, neuroinflammation as well as expression of SNAP-25, Akt, and pAkt were evaluated in the cerebral cortex. TA treatment protected against the impairment of memory in STZ-induced SDAT. STZ promoted an increase in neuronal death and the levels of proinflammatory cytokines (IL-6 and TNF-α) and a decrease in Akt and pAkt expression; TA was able to restore these changes. Neither STZ nor TA altered SNAP-25 expression or the levels of IL-12 and IL-4 in the cerebral cortex. Our study highlights that treatment with TA prevents memory deficits and reestablishes Akt and pAkt expression, protecting against neuronal death and neuroinflammation in STZ-induced SDAT in rats.
Subject(s)
Alzheimer Disease/metabolism , Inflammation Mediators/metabolism , Memory Disorders/metabolism , Proto-Oncogene Proteins c-akt/biosynthesis , Streptozocin/toxicity , Tannins/therapeutic use , Alzheimer Disease/chemically induced , Alzheimer Disease/prevention & control , Animals , Avoidance Learning/drug effects , Avoidance Learning/physiology , Cell Death/drug effects , Cell Death/physiology , Inflammation Mediators/antagonists & inhibitors , Male , Memory Disorders/chemically induced , Memory Disorders/prevention & control , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Rats , Rats, Wistar , Tannins/pharmacologyABSTRACT
Many aspects of the relationship between the demyelinating pathology and platelet function need to be elucidated. Thus, the activity of NTPDase and 5'-nucleotidase enzymes was analyzed in platelets from rats demyelinated with ethidium bromide (EB) and previously treated with ebselen (Ebs) and vitamin E (Vit. E). The animals were divided into four groups: for ebselen, the groups were: I-control (saline), II-(saline and Ebs), III-(EB) and IV-(EB and Ebs); and for vitamin E, the groups were: I - control (saline), II-(saline and Vit. E), III-(EB) and IV-(EB and Vit. E). After 3 and 21 days, the blood was collected and the platelets were separated for enzymatic assays. For the treatment with Ebs, the NTPDase activity for ATP substrate was significantly lower in groups II, III and IV (p < 0.05) after 3 days, while after 21 days, a reduction was observed in group III (p < 0.05). ADP hydrolysis was reduced in group II (p < 0.05) and increased in group IV (p < 0.05) after 3 days, while after 21 days there was an increase in group IV (p < 0.05). In the treatment with Vit. E, ATP hydrolysis was lower in groups II, III and IV (p < 0.05) after 3 and 21 days. ADP hydrolysis was increased in group II (p < 0.05) after 3 days, and in group IV (p < 0.05) after 21 days. However, 5'-nucleotidase activity was not altered by the treatments. These findings demonstrate that NTPDase activity in platelets is diminished in demyelinating events and the treatments with Ebs and Vit. E modulated adenine nucleotide hydrolysis.
Subject(s)
Adenine Nucleotides/metabolism , Antioxidants/pharmacology , Azoles/pharmacology , Blood Platelets/metabolism , Demyelinating Diseases/metabolism , Organoselenium Compounds/pharmacology , Vitamin E/pharmacology , 5'-Nucleotidase/metabolism , Adenosine Diphosphate/blood , Adenosine Monophosphate/blood , Adenosine Triphosphate/blood , Animals , Blood Platelets/drug effects , Demyelinating Diseases/chemically induced , Demyelinating Diseases/pathology , Ethidium , Hydrolysis , Isoindoles , Male , Pons/pathology , Rats , Rats, WistarABSTRACT
The activities of the enzymes NTPDase (EC 3.6.1.5, apyrase, CD39) and 5'-nucleotidase (EC 3.1.3.5, CD73) were analyzed in platelets from rats submitted to demyelination by ethidium bromide (EB) and treated with interferon beta (IFN-beta). The following groups were studied: I - control (saline), II - (saline and IFN-beta), III - (EB) and IV - (EB and IFN-beta). After 7, 15 and 30 days, the animals (n=7) were sacrificed and the platelets were separated by the method of Lunkes et al. [Lunkes, G., Lunkes D., Morsch, V., Mazzanti, C., Morsch, A., Miron, V., Schetinger, M.R.C., 2004. NTPDase and 5'-nucleotidase in rats alloxan- induced diabetes. Diabetes Research and Clinical Practice 65, 1-6]. NTPDase activity for ATP and ADP substrates was significantly lower in groups II and III after seven days, when compared to control (p<0.001). At fifteen days, ATP hydrolysis was significantly lower in group III and IV and higher in group II (p<0.001), while there was an activation of ADP hydrolysis in group II (p<0.001), when compared with the control. 5'-nucleotidase activity was significantly higher in group IV (p<0.001) after seven days, and lower in the groups III and IV (p<0.001) after fifteen days in relation to the control. No significant differences were observed in NTPDase and 5'-nucleotidase activities after thirty days. In conclusion, our study demonstrated that the hydrolysis of adenine nucleotides is modified in platelets of rats demyelinated and treated with IFN-beta.
Subject(s)
5'-Nucleotidase/metabolism , Adenine Nucleotides/metabolism , Antigens, CD/metabolism , Apyrase/metabolism , Blood Platelets , Demyelinating Diseases , Interferon-beta/therapeutic use , Animals , Blood Platelets/drug effects , Blood Platelets/enzymology , Blood Platelets/metabolism , Demyelinating Diseases/blood , Demyelinating Diseases/drug therapy , Demyelinating Diseases/enzymology , Disease Models, Animal , Ethidium , Hydrolysis , Male , Rats , Rats, WistarABSTRACT
OBJECTIVE: Here we investigated the impact of chronic high-intensity interval training (HIIT) and caffeine consumption on the activities of Na+-K+-ATPase and enzymes of the antioxidant system, as well as anxiolytic-like behaviour in the rat brain. METHODS: Animals were divided into groups: control, caffeine (4â mg/kg), caffeine (8â mg/kg), HIIT, HIIT plus caffeine (4â mg/kg) and HIIT plus caffeine (8â mg/kg). Rats were trained three times per week for 6 weeks, and caffeine was administered 30 minutes before training. We assessed the anxiolytic-like behaviour, Na+-K+-ATPase, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities, levels of reduced glutathione (GSH) and thiobarbituric acid reactive substances (TBARS) in the brain. RESULTS AND DISCUSSION: HIIT-induced anxiolytic-like behaviour increased Na+-K+-ATPase and GPx activities and TBARS levels, altered the activities of SOD and CAT in different brain regions, and decreased GSH levels. Caffeine, however, elicited anxiogenic-like behaviour and blocked HIIT effects. The combination of caffeine and HIIT prevented the increase in SOD activity in the cerebral cortex and GPx activity in three brain regions. Our results show that caffeine promoted anxiogenic behaviour and prevented HIIT-induced changes in the antioxidant system and Na+-K+-ATPase activities.
Subject(s)
Anti-Anxiety Agents/therapeutic use , Anxiety/drug therapy , Anxiety/metabolism , Caffeine/therapeutic use , Sodium-Potassium-Exchanging ATPase/metabolism , Animals , Antioxidants/metabolism , Catalase/metabolism , Glutathione/metabolism , Glutathione Peroxidase/metabolism , Lipid Peroxidation/drug effects , Male , Oxidative Stress/drug effects , Rats , Rats, Wistar , Superoxide Dismutase/metabolism , Thiobarbituric Acid Reactive Substances/metabolismABSTRACT
Peripheral inflammatory stimuli may activate a brain neuroinflammatory processes with consequences in brain function. The present study investigated if anthocyanins (ANT) consumption was able to prevent the memory loss, the neuronal damage, and the neuroinflammatory processes triggered by the intraperitoneal lipopolysaccharide (LPS) administration. C57BL6 male mice were treated with ANT (30-100 mg/kg by gavage). With a single dose or during 10 days, before be challenged with LPS (250 µg/kg intraperitoneally single administration), a classical inductor of inflammation. The data obtained showed that ANT was able to confer protection against the memory impairment after 10 days of ANT treatment (100 mg/kg). This phytonutrient also prevented the hypothermia episode induced by LPS. Moreover, ANT prevented the increase in protein carbonyl, NOx, and MDA levels in the hippocampus and cerebral cortex (4 and 24 h) in animal challenged with LPS. ANT showed a protective effect on the increase in the pro-inflammatory cytokines content, especially Interleukin (IL)-1ß, tumoral necrosis factor-α and on the reduction of IL-10 induced by LPS. ANT 100 mg/kg prevented the infiltration of peripheral immune cells in the hippocampus at 24 h post-LPS administration. In parallel, LPS increased the activity of myeloperoxidase in cortex and hippocampus, and ANT prevented this effect, also reducing microglia (Iba-1) and astrocyte (GFAP) immunoreactivity. Thus, our data support that ANT are a promising therapeutic component against brain disorders associated with process of neuroinflammation. Graphical Abstract á .
Subject(s)
Anthocyanins/therapeutic use , Inflammation/drug therapy , Memory Disorders/drug therapy , Animals , Anthocyanins/pharmacology , Cerebral Cortex/enzymology , Cerebral Cortex/pathology , Hippocampus/enzymology , Hippocampus/pathology , Hypothermia, Induced , Inflammation/complications , Inflammation Mediators/metabolism , Lipopolysaccharides/administration & dosage , Male , Memory Disorders/complications , Mice, Inbred C57BL , Models, Biological , Neuroglia/drug effects , Neuroglia/metabolism , Neuroglia/pathology , Neurons/drug effects , Neurons/metabolism , Neurons/pathology , Neuroprotective Agents/pharmacology , Neuroprotective Agents/therapeutic use , Oxidative Stress/drug effects , Peroxidase/metabolism , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
The use of ergogenic substances such as caffeine has become a strategy to enhance sports performance. In the present study we evaluated the effects of high-intensity interval training (HIIT) associated with caffeine intake on acetylcholinesterase (AChE) and Ca2+ATPase activity and glycogen levels in the muscles of rats were evaluated. The animals were divided in groups: control, caffeine 4 or 8mg/kg, HIIT, HIIT plus caffeine 4 or caffeine 8mg/kg. Our results showed a decrease in glycogen levels in muscle in all trained groups after acute session exercise, while that an increase in glycogen levels was observed in all groups in relation to control in chronic exercise protocol. HIIT increases the thickness of the left ventricle and the Ca2+-ATPase activity and decrease the AChE activity in gastrocnemius muscle. Caffeine treatment prevents changes in enzymes activities as well as left ventricular hypertrophy adaptation induced by HIIT. Our findings suggest that caffeine modulates crucial pathways for muscle contraction in HIIT.