Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 51
Filter
Add more filters











Publication year range
1.
Expert Opin Ther Targets ; 28(5): 401-418, 2024 May.
Article in English | MEDLINE | ID: mdl-38871633

ABSTRACT

INTRODUCTION: Inflammasome complexes, especially NLRP3, have gained great attention as a potential therapeutic target in mood disorders. NLRP3 triggers a caspase 1-dependent release of the inflammatory cytokines IL-1ß and IL-18, and seems to interact with purinergic and kynurenine pathways, all of which are implicated in mood disorders development and progression. AREAS COVERED: Emerging evidence supports NLRP3 inflammasome as a promising pharmacological target for mood disorders. We discussed the available evidence from animal models and human studies and provided a reflection on drawbacks and perspectives for this novel target. EXPERT OPINION: Several studies have supported the involvement of NLRP3 inflammasome in MDD. However, most of the evidence comes from animal models. The role of NLRP3 inflammasome in BD as well as its anti-manic properties is not very clear and requires further exploration. There is evidence of anti-manic effects of P2×R7 antagonists associated with reduction in the brain levels of IL-1ß and TNF-α in a murine model of mania. The involvement of other NLRP3 inflammasome expressing cells besides microglia, like astrocytes, and of other inflammasome complexes in mood disorders also deserves further investigation. Preclinical and clinical characterization of NLRP3 and other inflammasomes in mood disorders is needed before considering translational approaches, including clinical trials.


Subject(s)
Disease Models, Animal , Inflammasomes , Molecular Targeted Therapy , Mood Disorders , NLR Family, Pyrin Domain-Containing 3 Protein , Animals , Humans , Inflammasomes/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Mood Disorders/drug therapy , Mood Disorders/physiopathology , Mice , Bipolar Disorder/drug therapy , Bipolar Disorder/physiopathology , Purinergic P2X Receptor Antagonists/pharmacology , Purinergic P2X Receptor Antagonists/administration & dosage , Depressive Disorder, Major/drug therapy , Depressive Disorder, Major/physiopathology
2.
Braz J Med Biol Res ; 57: e13234, 2024.
Article in English | MEDLINE | ID: mdl-38716980

ABSTRACT

Patients undergoing chemotherapy with cisplatin commonly present gastrointestinal effects such as constipation and gastric emptying (GE) delay. Both the purinergic system and physical exercise modulate the gastrointestinal (GI) tract. In the current study, we investigated the role of ATP, physical exercise, and P2X7 receptor blocking on GE delay induced by cisplatin in rats. Male rats were divided into the following groups: control (C), cisplatin (Cis), exercise (Ex), Brilliant Blue G (BBG), ATP, Cis+Ex, Cis+ATP, Cis+BBG, Cis+Ex+BBG, Cis+Ex+BBG+ATP, and Cis+ATP+BBG. GE delay was induced by treatment with 1 mg/kg cisplatin (1 time/week for 5 weeks, ip). The moderate physical exercise was swimming (1 h/day, 5 days/week for 5 weeks). At the end of the treatment or exercise and 30 min before the GE assessment, some groups received BBG (50 mg/kg, sc) or ATP (2 mg/kg, sc). Then, GE was assessed after a 10-min postprandial period. Chronic use of Cis decreased GE delay (P<0.05) compared to the control group. Both exercise and ATP prevented (P<0.05) GE delay compared to Cis. The pretreatment with BBG significantly inhibited (P<0.05) the effect of exercise and ATP. On the other hand, the association between exercise and ATP reversed (P<0.05) the effect of the BBG and prevented GE delay. Therefore, we suggest that both exercise and treatment with ATP activate P2X7 receptors and prevent GE delay induced by cisplatin in rats.


Subject(s)
Adenosine Triphosphate , Antineoplastic Agents , Cisplatin , Gastric Emptying , Physical Conditioning, Animal , Rats, Wistar , Receptors, Purinergic P2X7 , Animals , Cisplatin/pharmacology , Male , Adenosine Triphosphate/metabolism , Gastric Emptying/drug effects , Gastric Emptying/physiology , Receptors, Purinergic P2X7/metabolism , Physical Conditioning, Animal/physiology , Antineoplastic Agents/pharmacology , Rats , Purinergic P2X Receptor Antagonists/pharmacology
3.
Braz J Med Biol Res ; 56: e12569, 2023.
Article in English | MEDLINE | ID: mdl-36856255

ABSTRACT

The purinergic system participates in the control of blood pressure. Hypertension promotes the occurrence of gastrointestinal disorders such as intestinal inflammation and gastric emptying delay. This study aimed i) to investigate the participation of the P2X7 receptor blocker Brilliant Blue G (BBG) on gastric emptying of solids and changes in oxidative stress in the gastric fundus, duodenum, and colon of spontaneously hypertensive rats (SHR) and ii) to study the putative relationship of this effect with the renin-angiotensin system. Rats were divided into five groups: Control, SHR, SHR+BBG, SHR+BBG+ATP, and SHR+BBG+ANG II. In the gastrointestinal tract, we assessed gastric emptying (GE) and oxidative stress markers (NOx, MPO, GSH, SOD). We observed a decrease in the GE rate (P<0.05) in SHR vs control rats (21.8±2.0% vs 42.8±3.5%). The decrease in GE was returned (P<0.05) to control levels by BBG in SHR rats (21.8±2.0% vs 41.6±3.2%). Co-administration of ATP or ANG II together with BBG bypassed the effect of the P2X7 antagonist on GE in SHR (P<0.05) (21.9±5.0% vs 25.6±3.0% vs 41.6±3.2%). The MPO activity increased (P<0.05) in the gastric fundus of SHR compared to control rats (6.12±2.26 vs 0.077±0.02 UMPO/mg tissue); this effect was prevented (P<0.05) by BBG (0.55±0.15 vs 6.12±2.26 UMPO/mg tissue). Data demonstrated that blockage of P2X7 receptors with BBG can improve the GE delay and oxidative stress biomarkers in SHR animals. This preventive effect of BBG on GE delay was abrogated by ANG II and ATP, thus prompting crosstalk between renin-angiotensin and the purinergic signaling systems underlying this phenomenon.


Subject(s)
Gastrointestinal Diseases , Purinergic P2X Receptor Antagonists , Rats , Animals , Rats, Inbred SHR , Purinergic P2X Receptor Antagonists/pharmacology , Receptors, Purinergic P2X7 , Adenosine Triphosphate
4.
Molecules ; 28(2)2023 Jan 06.
Article in English | MEDLINE | ID: mdl-36677652

ABSTRACT

ATP acts in the extracellular environment as an important signal, activating a family of receptors called purinergic receptors. In recent years, interest in the potential therapeutics of purinergic components, including agonists and antagonists of receptors, has increased. Currently, many observations have indicated that ATP acts as an important mediator of inflammatory responses and, when found in high concentrations in the extracellular space, is related to the activation of the P2X7 purinergic receptor. In this sense, the search for new inhibitors for this receptor has attracted a great deal of attention in recent years. Sulfonamide derivatives have been reported to be potent inhibitors of P2X receptors. In this study, ten naphthoquinone sulfonamide derivatives and five naphthoquinone sulfonate ester derivatives were tested for their inhibitory activity on the P2X7 receptor expressed in peritoneal macrophages. Some compounds showed promising results, displaying IC50 values lower than that of A740003. Molecular docking and dynamic studies also indicated that the active compounds bind to an allosteric site on P2X7R. The binding free energy indicates that sulfonamides have an affinity for the P2X7 receptor similar to A740003. Therefore, the compounds studied herein present potential P2X7R inhibition.


Subject(s)
Naphthoquinones , Purinergic P2X Receptor Antagonists , Purinergic P2X Receptor Antagonists/pharmacology , Purinergic P2X Receptor Antagonists/chemistry , Sulfonamides/pharmacology , Molecular Docking Simulation , Naphthoquinones/pharmacology , Naphthoquinones/chemistry , Receptors, Purinergic P2X7 , Adenosine Triphosphate/metabolism
5.
J Bioenerg Biomembr ; 54(5-6): 227-239, 2022 12.
Article in English | MEDLINE | ID: mdl-36070071

ABSTRACT

The P2X7 receptor (P2X7R) is an ion channel that promotes the passage of ions through the membrane through brief stimulation once activated by ATP, its endogenous opener. However, prolonged stimulation with ATP, which occurs in pathological processes, opens a nonselective pore in the plasma membrane, allowing the passage of large molecules and leading to cytokine release or even cell death. In this sense, the search for new inhibitors for this receptor has attracted a great deal of attention in recent years. Considering the booming of biomass upgrading reactions in recent years and the continued efforts to synthesize biologically active molecules containing the 1,2,3-triazole ring, in the present work, we aimed to investigate whether triazole-linked menadione-furan derivatives could present P2X7R inhibitory activity. The novel compounds were tested for their inhibitory activity on ATP-induced dye uptake in peritoneal macrophages. Some have shown promising results, having displayed IC50 values lower than that of the P2X7R inhibitor BBG. Molecular docking studies also indicated that the active compounds bind to an allosteric site on P2X7R, presenting potential P2X7R inhibition.


Subject(s)
Triazoles , Vitamin K 3 , Molecular Docking Simulation , Triazoles/pharmacology , Adenosine Triphosphate/pharmacology , Furans/pharmacology , Receptors, Purinergic P2X7 , Purinergic P2X Receptor Antagonists/pharmacology
6.
World J Gastroenterol ; 28(30): 4075-4088, 2022 Aug 14.
Article in English | MEDLINE | ID: mdl-36157120

ABSTRACT

BACKGROUND: Clostridioides difficile (C. difficile) is the most common pathogen causing health care-associated infections. C. difficile TcdA and TcdB have been shown to activate enteric neurons; however, what population of these cells is more profoundly influenced and the mechanism underlying these effects remain unknown. AIM: To characterize a specific population of TcdA-affected myenteric neurons and investigate the role of the P2X7 receptor in TcdA-induced ileal inflammation, cell death, and the changes in the enteric nervous system in mice. METHODS: Swiss mice were used to model TcdA-induced ileitis in ileal loops exposed to TcdA (50 µg/Loop) for 4 h. To investigate the role of the P2X7 receptor, Brilliant Blue G (50 mg/kg, i.p.), which is a nonspecific P2X7 receptor antagonist, or A438079 (0.7 µg/mouse, i.p.), which is a competitive P2X7 receptor antagonist, were injected one hour prior to TcdA challenge. Ileal samples were collected to analyze the expression of the P2X7 receptor (by quantitative real-time polymerase chain reaction and immunohistochemistry), the population of myenteric enteric neurons (immunofluorescence), histological damage, intestinal inflammation, cell death (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling), neuronal loss, and S100B synthesis (immunohistochemistry). RESULTS: TcdA upregulated (P < 0.05) the expression of the P2X7 receptor gene in the ileal tissues, increasing the level of this receptor in myenteric neurons compared to that in control mice. Comparison with the control mice indicated that TcdA promoted (P < 0.05) the loss of myenteric calretinin+ (Calr) and choline acetyltransferase+ neurons and increased the number of nitrergic+ and Calr+ neurons expressing the P2X7 receptor. Blockade of the P2X7 receptor decreased TcdA-induced intestinal damage, cytokine release [interleukin (IL)-1ß, IL-6, IL-8, and tumor necrosis factor-α], cell death, enteric neuron loss, and S100B synthesis in the mouse ileum. CONCLUSION: Our findings demonstrated that TcdA induced the upregulation of the P2X7 receptor, which promoted enteric neuron loss, S100B synthesis, tissue damage, inflammation, and cell death in the mouse ileum. These findings contribute to the future directions in understanding the mechanism involved in intestinal dysfunction reported in patients after C. difficile infection.


Subject(s)
Bacterial Toxins , Clostridioides difficile , Ileitis , Animals , Apoptosis , Biotin/metabolism , Calbindin 2 , Choline O-Acetyltransferase/metabolism , DNA Nucleotidylexotransferase/metabolism , Enterotoxins , Ileitis/chemically induced , Inflammation/pathology , Interleukin-6/metabolism , Interleukin-8/metabolism , Mice , Neurons/pathology , Purinergic P2X Receptor Antagonists/pharmacology , Receptors, Purinergic P2X7 , Tumor Necrosis Factor-alpha/metabolism
7.
Front Immunol ; 13: 872760, 2022.
Article in English | MEDLINE | ID: mdl-36032155

ABSTRACT

Previous studies have demonstrated that acute colonic inflammation leads to an increase in dorsal root ganglia (DRG) neuronal excitability. However, the signaling elements implicated in this hyperexcitability have yet to be fully unraveled. Extracellular adenosine 5'-triphosphate (ATP) is a well-recognized sensory signaling molecule that enhances the nociceptive response after inflammation through activation of P2X3 receptors, which are expressed mainly by peripheral sensory neurons. The aim of this study is to continue investigating how P2X3 affects neuronal hypersensitivity in an acute colitis animal model. To achieve this, DNBS (Dinitrobenzene sulfonic acid; 200 mg/kg) was intrarectally administered to C57BL/6 mice, and inflammation severity was assessed according to the following parameters: weight loss, macroscopic and microscopic scores. Perforated patch clamp technique was used to evaluate neuronal excitability via measuring changes in rheobase and action potential firing in T8-L1 DRG neurons. A-317491, a well-established potent and selective P2X3 receptor antagonist, served to dissect their contribution to recorded responses. Protein expression of P2X3 receptors in DRG was evaluated by western blotting and immunofluorescence. Four days post-DNBS administration, colons were processed for histological analyses of ulceration, crypt morphology, goblet cell density, and immune cell infiltration. DRG neurons from DNBS-treated mice were significantly more excitable compared with controls; these changes correlated with increased P2X3 receptor expression. Furthermore, TNF-α mRNA expression was also significantly higher in inflamed colons compared to controls. Incubation of control DRG neurons with TNF-α resulted in similar cell hyperexcitability as measured in DNBS-derived neurons. The selective P2X3 receptor antagonist, A-317491, blocked the TNF-α-induced effect. These results support the hypothesis that TNF-α enhances colon-innervating DRG neuron excitability via modulation of P2X3 receptor activity.


Subject(s)
Colitis , Ganglia, Spinal , Adenosine Triphosphate , Animals , Inflammation , Mice , Mice, Inbred C57BL , Purinergic P2X Receptor Antagonists , Receptors, Purinergic P2X3 , Sensory Receptor Cells , Tumor Necrosis Factor-alpha
8.
Behav Brain Res ; 423: 113786, 2022 04 09.
Article in English | MEDLINE | ID: mdl-35124136

ABSTRACT

The zebrafish has been considered an ideal model for studies of complex behaviors since its behavioral repertoire is well described. Therefore, this study evaluated the perceived pain through behavioral changes in zebrafish larvae. Here we investigated the Acetic Acid (AA) effects on zebrafish larvae exposed in a short-time period (60 s) and the preventive effect from routinely used compounds, Dimethyl Sulfoxide (DMSO), Ethanol (EtOH), Ibuprofen (IBP), and Paracetamol (PAR). In addition, the effect of P2×7 antagonist, A740003, and pannexin channel 1 (PANX-1) inhibitor Probenecid (PROB) on AA-induced behavioral changes were evaluated. AA impaired the distance covered, acceleration, movement, and latency to the first entry in the center from 5 dpf exposed larvae. At 0.050% AA, PAR prevented alterations from the distance covered, acceleration, and movement. Surprisingly, 0.3% DMSO prevented behavioral changes induced by AA. However, the effects from 0.2% DMSO were not prominent. We used 0.2% DMSO as a PROB diluent. PROB prevented the changes in distance and movement observed at both AA concentrations (0.0025% and 0.05%) tested. Since EtOH had no analgesic properties, we used it as an A740003 vehicle to observe the analgesic effects of this compound. As noted, A740003 did not prevent the behavioral changes in the AA-induced pain model. In contrast, 0.2% DMSO and PROB prevented AA-induced behavioral changes. These data enforce that zebrafish could be used in translational studies since this species has behavioral responses related to pain in the early stages of development and responses to analgesics similar to observed in mammals.


Subject(s)
Analgesics/pharmacology , Behavior, Animal/drug effects , Connexins , Dimethyl Sulfoxide/pharmacology , Pain , Purinergic P2X Receptor Antagonists/pharmacology , Receptors, Purinergic P2X7 , Zebrafish Proteins , Animals , Connexins/antagonists & inhibitors , Connexins/metabolism , Disease Models, Animal , Larva , Pain/drug therapy , Pain/metabolism , Receptors, Purinergic P2X7/metabolism , Zebrafish , Zebrafish Proteins/antagonists & inhibitors , Zebrafish Proteins/metabolism
9.
Histochem Cell Biol ; 157(1): 65-81, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34626216

ABSTRACT

Inflammatory bowel diseases (IBDs) are chronic diseases of the gastrointestinal tract that include ulcerative colitis and Crohn's disease and affect enteric neurons. Research has shown that Brilliant Blue G (BBG), a P2X7 receptor antagonist, restores enteric neurons following ischemia and reperfusion. This study aimed to evaluate the effect of BBG on myenteric neurons of the distal colon in an experimental rat model of ulcerative colitis. Colitis was induced by injection of 2,4,6-trinitrobenzene sulfonic acid (TNBS) into the large intestine. BBG was administered 1 h after colitis induction and for five consecutive days thereafter. Distal colons were collected 24 h or 7 days after TNBS injection. The animals were divided into 24-h and 7-day sham (vehicle injection rather than colitis induction), 24-h colitis, 24-h BBG, 7-day colitis and 7-day BBG groups. The disease activity index (DAI), neuronal density and profile of neuronal nitric oxide synthase (nNOS)-, choline acetyltransferase (ChAT)- and P2X7 receptor-immunoreactive enteric neurons were analyzed, and histological analysis was performed. The results showed recovery of the DAI and histological tissue integrity in the BBG groups compared to those in the colitis groups. In addition, the numbers of neurons positive for nNOS, ChAT and the P2X7 receptor per area were decreased in the colitis groups, and these measures were recovered in the BBG groups. Neuronal size was increased in the colitis groups and restored in the BBG groups. In conclusion, BBG is effective in improving experimental ulcerative colitis, and the P2X7 receptor may be a therapeutic target.


Subject(s)
Colitis, Ulcerative , Animals , Colitis, Ulcerative/chemically induced , Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/pathology , Colon/pathology , Neurons/pathology , Purinergic P2X Receptor Antagonists/pharmacology , Rats , Receptors, Purinergic P2X7
10.
Biomed Pharmacother ; 142: 112006, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34392085

ABSTRACT

P2X7 receptor promotes inflammatory response and neuropathic pain. New drugs capable of impairing inflammation and pain-reducing adverse effects extracted from plant extracts have been studied. Physalis angulate L. possesses traditional uses and exhibits antiparasitic, anti-inflammatory, antimicrobial, antinociceptive, antimalarial, antileishmanial, immunosuppressive, antiasthmatic. diuretic, and antitumor activities. The most representative phytochemical constituents identified with medicinal importance are the physalins and withanolides. However, the mechanism of anti-inflammatory action is scarce. Although some physalins and withanolides subtypes have anti-inflammatory activity, only four physalins subtypes (B, D, F, and G) have further studies. Therefore, we evaluated the crude ethanolic extract enriched with physalins B, D, F, and G from P. angulata leaves, a pool containing the physalins B, D, F, G, and the physalins individually, as P2X7 receptor antagonists. For this purpose, we evaluated ATP-induced dye uptake, macroscopic currents, and interleukin 1-ß (IL-1ß) in vitro. The crude extract and pool dose-dependently inhibited P2X7 receptor function. Thus, physalin B, D, F, and G individually evaluated for 5'-triphosphate (ATP)-induced dye uptake assay, whole-cell patch-clamp, and cytokine release showed distinct antagonist levels. Physalin D displayed higher potency and efficacy than physalin B, F, and G for all these parameters. In vivo mice model as ATP-induced paw edema was potently inhibited for physalin D, in contrast to physalin B, F, and G. ATP and lipopolysaccharide (LPS)-induced pleurisy in mice were reversed for physalin D treatment. Molecular modeling and computational simulation predicted the intermolecular interactions between the P2X7 receptor and physalin derivatives. In silico results indicated physalin D and F as a potent allosteric P2X7 receptor antagonist. These data confirm physalin D as a promisor source for developing a new P2X7 receptor antagonist with anti-inflammatory action.


Subject(s)
Acute Lung Injury/drug therapy , Physalis/chemistry , Plant Extracts/pharmacology , Secosteroids/pharmacology , Acute Lung Injury/physiopathology , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/isolation & purification , Anti-Inflammatory Agents/pharmacology , Computer Simulation , Disease Models, Animal , Dose-Response Relationship, Drug , HEK293 Cells , Humans , Male , Mice , Models, Molecular , Plant Extracts/administration & dosage , Plant Leaves , Purinergic P2X Receptor Antagonists/administration & dosage , Purinergic P2X Receptor Antagonists/isolation & purification , Purinergic P2X Receptor Antagonists/pharmacology , Receptors, Purinergic P2X7/drug effects , Secosteroids/isolation & purification
11.
Purinergic Signal ; 16(3): 403-414, 2020 09.
Article in English | MEDLINE | ID: mdl-32766958

ABSTRACT

This study aimed to evaluate whether the development and/or maintenance of chronic-latent muscle hyperalgesia is modulated by P2X3 receptors. We also evaluate the expression of P2X3 receptors and PKCε of dorsal root ganglions during these processes. A mouse model of chronic-latent muscle hyperalgesia, induced by carrageenan and evidenced by PGE2, was used. Mechanical muscle hyperalgesia was measured by Randall-Selitto analgesimeter. The involvement of P2X3 receptors was analyzed by using the selective P2X3 receptors antagonist A-317491 by intramuscular or intrathecal injections. Expression of P2X3 and PKCε in dorsal root ganglion (L4-S1) were evaluated by Western blotting. Intrathecal blockade of P2X3 receptors previously to carrageenan prevented the development and maintenance of acute and chronic-latent muscle hyperalgesia, while intramuscular blockade of P2X3 receptors previously to carrageenan only reduced the acute muscle hyperalgesia and had no effect on chronic-latent muscle hyperalgesia. Intrathecal, but not intramuscular, blockade of P2X3 receptors immediately before PGE2, in animals previously sensitized by carrageenan, reversed the chronic-latent muscle hyperalgesia. There was an increase in total and phosphorylated PKCε 48 h after the beginning of acute muscle hyperalgesia, and in P2X3 receptors at the period of chronic muscle hyperalgesia. P2X3 receptors expressed on spinal cord dorsal horn contribute to transition from acute to chronic muscle pain. We also suggest an interaction of PKCε and P2X3 receptors in this process. Therefore, we point out P2X3 receptors of the spinal cord dorsal horn as a pharmacological target to prevent the development or reverse the chronic muscle pain conditions.


Subject(s)
Acute Pain/metabolism , Chronic Pain/metabolism , Muscle, Skeletal/metabolism , Myalgia/metabolism , Receptors, Purinergic P2X3/metabolism , Animals , Disease Progression , Ganglia, Spinal/drug effects , Ganglia, Spinal/metabolism , Male , Mice , Muscle, Skeletal/drug effects , Pain Threshold/drug effects , Phenols/pharmacology , Polycyclic Compounds/pharmacology , Purinergic P2X Receptor Antagonists/pharmacology
12.
BMC Nephrol ; 21(1): 206, 2020 05 29.
Article in English | MEDLINE | ID: mdl-32471386

ABSTRACT

BACKGROUND: Previous study showed that purinergic P2X7 receptors (P2X7R) reach the highest expression in the first week after unilateral ureteral obstruction (UUO) in mice, and are involved in the process of inflammation, apoptosis and fibrosis of renal tissue. We, herein, document the role of purinergic P2X7 receptors activation on the third day of UUO, as assessed by means of BBG as its selective inhibitor. METHODS: We investigated the effects of brilliant blue G (BBG), a P2X7R antagonist, in the third day of kidney tissue response to UUO in rats. For this purpose, male Wistar rats submitted to UUO or sham operated, received BBG or vehicle (V), comprising four groups: UUO-BBG, UUO-V, sham-BBG and sham-V. The kidneys were harvested on day 3 UUO and prepared for histology, immunohistochemistry (P2X7R, PCNA, CD-68, α-sma, TGF-ß1, Heat-shock protein-47, TUNEL assay), quantitative real-time PCR (IL-1ß, procollagens type I, III, and IV) for mRNA quantification. RESULTS: The group UUO-V presented an enhancement in tubular cell P2X7-R expression, increase influx of macrophages and myofibroblasts, HSP-47 and TGF- ß1 expression. Also, upregulation of procollagen types I, III, and IV, and IL-1ß mRNAs were seen. On the other hand, group UUO-BBG showed lower expression of procollagens and IL-1ß mRNAs, as well as less immunoreactivity of HSP-47, TGF-ß, macrophages, myofibroblasts, and tubular apoptosis. This group also presented increased epithelial cell proliferation. CONCLUSION: BBG, a known highly selective inhibitor of P2X7R, attenuated renal inflammation, collagen synthesis, renal cell apoptosis, and enhanced renal cell proliferation in the early phase of rat model of UUO.


Subject(s)
Cell Proliferation/drug effects , Kidney/pathology , Nephritis/drug therapy , Purinergic P2X Receptor Antagonists/therapeutic use , Rosaniline Dyes/therapeutic use , Ureteral Obstruction/complications , Actins/metabolism , Animals , Antigens, CD/metabolism , Antigens, Differentiation, Myelomonocytic/metabolism , Apoptosis/drug effects , Cell Movement , Collagen Type I/genetics , Collagen Type III/genetics , Collagen Type IV/genetics , Fibrosis , HSP47 Heat-Shock Proteins/metabolism , Interleukin-1beta/genetics , Kidney/metabolism , Kidney Tubules/pathology , Macrophages/physiology , Male , Myofibroblasts/physiology , Nephritis/etiology , Purinergic P2X Receptor Antagonists/pharmacology , RNA, Messenger/metabolism , Rats , Rats, Wistar , Rosaniline Dyes/pharmacology , Time Factors , Transforming Growth Factor beta1/metabolism , Up-Regulation
13.
Respir Physiol Neurobiol ; 277: 103438, 2020 06.
Article in English | MEDLINE | ID: mdl-32259688

ABSTRACT

Hyperreflexia of the peripheral chemoreceptors is a potential contributor of apnoeas of prematurity (AoP). Recently, it was shown that elevated P2X3 receptor expression was associated with elevated carotid body afferent sensitivity. Therefore, we tested whether P2X3 receptor antagonism would reduce AoP known to occur in newborn rats. Unrestrained whole-body plethysmography was used to record breathing and from this the frequency of apnoeas at baseline and following administration of either a P2X3 receptor antagonist - AF-454 (5 mg/kg or 10 mg/kg s.c.) or vehicle was derived. In a separate group, we tested the effects of AF-454 (10 mg/kg) on the hypoxic ventilatory response (10 % FiO2). Ten but not 5 mg/kg AF-454 reduced the frequency of AoP and improved breathing regularity significantly compared to vehicle. Neither AF-454 (both 5 and 10 mg/kg) nor vehicle affected baseline respiration. However, P2X3 receptor antagonism (10 mg/kg) powerfully blunted hypoxic ventilatory response to 10 % FiO2. These data suggest that P2X3 receptors contribute to AoP and the hypoxic ventilatory response in newborn rats but play no role in the drive to breathe at rest.


Subject(s)
Apnea/prevention & control , Purinergic P2X Receptor Antagonists/therapeutic use , Receptors, Purinergic P2X3/physiology , Animals , Animals, Newborn , Apnea/physiopathology , Carotid Body/drug effects , Carotid Body/physiopathology , Hypoxia/drug therapy , Hypoxia/physiopathology , Male , Plethysmography, Whole Body/methods , Purinergic P2X Receptor Antagonists/pharmacology , Rats , Rats, Wistar
14.
Mol Neurobiol ; 57(3): 1347-1360, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31729632

ABSTRACT

The pathophysiology of bipolar disorder remains incompletely elucidated. The purinergic receptor, P2X7 (P2X7R), plays a central role in neuroinflammation, the establishment, and maintenance of microglial activation and neuronal damage/death, all characteristics of bipolar disorder pathology. The present study aims to explore the participation of the P2X7R in a preclinical pharmacological model of mania. We analyzed the modulatory effects of the P2X7R antagonist, brilliant blue, on behavior, monoamines, gene expression, serum purine levels, and cell typing in a pharmacological model of mania induced by D-amphetamine (AMPH) in mice. Our results corroborate an association between the P2X7 receptor and the preclinical animal model of mania, as demonstrated by the decreased responsiveness to AMPH in animals with pharmacologically blocked P2X7R. This study further suggests a possible dopaminergic mechanism for the action of P2X7 receptor antagonism. Additionally, we observed increased peripheral levels of adenosine, a neuroprotective molecule, and increased central expression of Entpd3 and Entpd1 leading to the hydrolysis of ATP, a danger signal, possibly as an attempt to compensate for the damage induced by AMPH. Lastly, P2X7R antagonism in the AMPH model was found to potentially modulate astrogliosis. Our results support the hypothesis that P2X7R plays a vital role in the pathophysiology of mania, possibly by modulating the dopaminergic pathway and astrogliosis, as reflected in the behavioral changes observed. Taken together, this study suggests that a purinergic system imbalance is associated with the AMPH-induced preclinical animal model of mania. P2X7R may represent a promising molecular therapeutic target for bipolar disorder.


Subject(s)
Bipolar Disorder/physiopathology , Hippocampus/drug effects , Purinergic P2X Receptor Antagonists/pharmacology , Receptors, Purinergic P2X7/drug effects , Adenosine Triphosphate/metabolism , Animals , Bipolar Disorder/drug therapy , Bipolar Disorder/metabolism , Cell Death/drug effects , Disease Models, Animal , Gliosis/drug therapy , Hippocampus/metabolism , Hippocampus/pathology , Male , Mice, Inbred C57BL , Receptors, Purinergic P2X7/metabolism
15.
J Psychopharmacol ; 33(11): 1436-1446, 2019 11.
Article in English | MEDLINE | ID: mdl-31526216

ABSTRACT

BACKGROUND: P2X7 receptors (P2X7R) are ligand-gated ion channels activated by adenosine 5'-triphosphate (ATP), which are involved in processes that are dysfunctional in stress response and depression, such as neurotransmitter release, and neuroimmune response. Genetic and pharmacological inhibition of the P2X7R induce antidepressant-like effects in animals exposed to stress. However, the effect of P2X7R antagonism in an animal model of depression based on selective breeding has not previously been studied, and the mechanism underling the antidepressant-like effect induced by the P2X7R blockade remains unknown. AIMS: The present study aimed to: (1) determine whether P2X7R blockade induces antidepressant-like effects in the Flinders Sensitive Line (FSL) rats and, (2) investigate whether brain-derived neurotrophic factor (BDNF) signalling in the frontal cortex and hippocampus is involved in this effect. METHODS: FSL and the control Flinders Resistant Line (FRL) rats were treated with vehicle or the P2X7R antagonist A-804598 (3, 10 or 30 mg/Kg/day) for 1 or 7 days before being exposed to the forced swim test (FST). After the behavioural test, animals were decapitated, their brains were removed and the frontal cortex, ventral and dorsal hippocampus were dissected for BDNF signalling analysis. RESULTS: We found that repeated treatment with A-804598 (30 mg/Kg) reduced the immobility time in the FST and activated the BDNF signalling in the ventral hippocampus of FSL rats. CONCLUSIONS: P2X7R blockade induces an antidepressant-like effect associated with increased levels of BDNF-AKT-p70 S6 kinase in the ventral hippocampus, which may be mediated by tropomyosin-related kinase B (TRKB) receptor activation supporting the notion of P2X7R antagonism as a potential new antidepressant strategy.


Subject(s)
Antidepressive Agents/pharmacology , Depression/drug therapy , Guanidines/pharmacology , Purinergic P2X Receptor Antagonists/administration & dosage , Quinolines/pharmacology , Animals , Antidepressive Agents/administration & dosage , Behavior, Animal/drug effects , Brain-Derived Neurotrophic Factor/metabolism , Depression/physiopathology , Disease Models, Animal , Guanidines/administration & dosage , Male , Maximum Tolerated Dose , Purinergic P2X Receptor Antagonists/pharmacology , Quinolines/administration & dosage , Rats , Receptor, trkB/metabolism , Receptors, Purinergic P2X7/drug effects , Receptors, Purinergic P2X7/metabolism , Stress, Psychological/drug therapy , Swimming , Time Factors
16.
J Bioenerg Biomembr ; 51(4): 277-290, 2019 08.
Article in English | MEDLINE | ID: mdl-31256283

ABSTRACT

The P2X7 receptor (P2X7R) is an ion channel which is activated by interactions with the extracellular ATP molecules. The molecular complex P2X7R/ATP induces conformational changes in the protein subunits, opening a pore in the ion channel macromolecular structure. Currently, the P2X7R has been studied as a potential therapeutic target of anti-inflammatory drugs. Based on this, a series of eight boronic acids (NO) analogs were evaluated on the biologic effect of this pharmacophoric group on the human and murine P2X7R. The boronic acids derivatives NO-01 and NO-12 inhibited in vitro human and murine P2X7R function. These analogs compounds showed effect better than compound BBG and similar to inhibitor A740003 for inhibiting dye uptake, in vitro IL-1ß release and ATP-induced paw edema in vivo. In both, in vitro and in vivo assays the compound NO-01 showed to be the hit compound in the present series of the arylboronic acids analogs. The molecular docking suggests that the NO derivatives bind into the upper body domain of the P2X7 pore and that the main intermolecular interaction with the two most active NO derivatives occur with the residues Phe 95, 103 and 293 by hydrophobic interactions and with Leu97, Gln98 and Ser101 by hydrogen bonds.. These results indicate that the boronic acid derivative NO-01 shows the lead compound characteristics to be used as a scaffold structure to the development of new P2X7R inhibitors with anti-inflammatory action.


Subject(s)
Anti-Inflammatory Agents , Boronic Acids , Purinergic P2X Receptor Antagonists , Receptors, Purinergic P2X7/metabolism , Acetamides/chemistry , Acetamides/pharmacology , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Boronic Acids/chemistry , Boronic Acids/pharmacology , HEK293 Cells , Humans , Inflammation/drug therapy , Inflammation/genetics , Inflammation/metabolism , Inflammation/pathology , Male , Mice , Purinergic P2X Receptor Antagonists/chemistry , Purinergic P2X Receptor Antagonists/pharmacology , Quinolines/chemistry , Quinolines/pharmacology , Receptors, Purinergic P2X7/genetics
17.
Article in English | MEDLINE | ID: mdl-31136852

ABSTRACT

Copper is a metal that participates in several essential reactions in living organisms, and it has been used as an inflammatory inducing agent in zebrafish larvae. In this study, we evaluated the effect P2X7 receptor and/or pannexin channel 1 (PANX-1) blockage in this inflammation model. To perform the experiments, 7 dpf larvae were exposed to 10 µM of copper and treated with 100 µM probenecid, PANX-1 inhibitor, and/or 300 nM A740003, a P2X7R selective antagonist. Larvae survival was assessed up to 24 h after treatments. The evaluation of larvae behavior was evaluated after acute (4 h) and chronic (24 h) exposure. The parameters of locomotor activity measured were: mobile time, average speed, distance and turn angle. We analyzed the gene expression of the P2X7 receptor, PANX1a and PANX1b channels and interleukins IL-10 and IL-1b after 24 h of treatment. Treatments did not decrease larval survival in the time interval studied. Changes in larvae locomotion were observed after the longest time of exposure to copper and the treatment with probenecid was able to reverse part of the effects caused by copper. No significant difference was observed in the oxidative stress assays and probenecid and copper treatment decrease partially PANX1a gene expression groups. The data presented herein shows the relevance of the blockage of P2X7-PANX-1 in copper-induced inflammation.


Subject(s)
Connexins/genetics , Copper/toxicity , Inflammation/chemically induced , Receptors, Purinergic P2X7/genetics , Zebrafish Proteins/genetics , Zebrafish/metabolism , Acetamides/pharmacology , Animals , Connexins/antagonists & inhibitors , Disease Models, Animal , Female , Gene Expression Regulation/drug effects , Inflammation/mortality , Interleukin-10/genetics , Interleukin-1beta/genetics , Larva/drug effects , Locomotion/drug effects , Male , Oxidative Stress , Probenecid/pharmacology , Purinergic P2X Receptor Antagonists/toxicity , Quinolines/pharmacology , Zebrafish/genetics , Zebrafish Proteins/antagonists & inhibitors
18.
Purinergic Signal ; 15(2): 167-175, 2019 06.
Article in English | MEDLINE | ID: mdl-31115830

ABSTRACT

P2X3 receptors are involved with several pain conditions. Muscle pain induced by static contraction has an important socioeconomic impact. Here, we evaluated the involvement of P2X3 receptors on mechanical muscle hyperalgesia and neutrophil migration induced by static contraction in rats. Also, we evaluated whether static contraction would be able to increase muscle levels of TNF-α and IL-1ß. Male Wistar rats were pretreated with the selective P2X3 receptor antagonist, A-317491, by intramuscular or intrathecal injection and the static contraction-induced mechanical muscle hyperalgesia was evaluated using the Randall-Selitto test. Neutrophil migration was evaluated by measurement of myeloperoxidase (MPO) kinetic-colorimetric assay and the cytokines TNF-α and IL-1ß by enzyme-linked immunosorbent assay. Intramuscular or intrathecal pretreatment with A-317491 prevented static contraction-induced mechanical muscle hyperalgesia. In addition, A-317491 reduced static contraction-induced mechanical muscle hyperalgesia when administered 30 and 60 min of the beginning of static contraction, but not after 30 and 60 min of the end of static contraction. Intramuscular A-317491 also prevented static contraction-induced neutrophil migration. In a period of 24 h, static contraction did not increase muscle levels of TNF-α and IL-1ß. These findings demonstrated that mechanical muscle hyperalgesia and neutrophil migration induced by static contraction are modulated by P2X3 receptors expressed on the gastrocnemius muscle and spinal cord dorsal horn. Also, we suggest that P2X3 receptors are important to the development but not to maintenance of muscle hyperalgesia. Therefore, P2X3 receptors can be pointed out as a target to musculoskeletal pain conditions induced by daily or work-related activities.


Subject(s)
Myalgia/metabolism , Neutrophils , Receptors, Purinergic P2X3/metabolism , Animals , Cell Movement , Hyperalgesia/etiology , Hyperalgesia/metabolism , Male , Muscle Contraction/drug effects , Muscle Contraction/physiology , Myalgia/etiology , Neutrophils/drug effects , Phenols/pharmacology , Polycyclic Compounds/pharmacology , Purinergic P2X Receptor Antagonists/pharmacology , Rats , Rats, Wistar
19.
Acta Diabetol ; 56(9): 1031-1036, 2019 Sep.
Article in English | MEDLINE | ID: mdl-30982154

ABSTRACT

AIMS: Retinopathy is a leading cause of vision impairment in diabetes. Its pathogenesis involves inflammation, pathological angiogenesis, neuronal and glial dysfunction. The purinergic P2X7 receptor (P2X7R) has a leading role in inflammation and angiogenesis. Potent and selective P2X7R blockers have been synthesized and tested in Phase I/II clinical studies. We hypothesize that P2X7R blockade will ameliorate diabetes-related pathological retinal changes. METHODS: Streptozotocin (STZ)-treated rats were intraperitoneally inoculated with either of two small molecule P2X7R receptor inhibitors, A740003 and AZ10606120, and after blood glucose levels increased to above 400 mg/dL, retinae were analyzed for P2X7R expression, vascular permeability, VEGF, and IL-6 expression. RESULTS: STZ administration caused a near fourfold increase in blood glucose, a large increase in retinal microvasculature permeability, as well as in retinal P2X7R, VEGF, and IL-6 expression. P2X7R blockade fully reversed retinal vascular permeability increase, VEGF accumulation, and IL-6 expression, with no effect on blood glucose. CONCLUSION: P2X7R blockade might be promising strategy for the treatment of microvascular changes observed in the early phases of diabetic retinopathy.


Subject(s)
Diabetes Mellitus, Type 1/complications , Diabetes Mellitus, Type 1/drug therapy , Diabetic Retinopathy/prevention & control , Purinergic P2X Receptor Antagonists/pharmacology , Retina/drug effects , Animals , Blood Glucose/drug effects , Blood Glucose/metabolism , Diabetes Mellitus, Experimental/complications , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Experimental/pathology , Diabetes Mellitus, Type 1/pathology , Diabetic Retinopathy/drug therapy , Diabetic Retinopathy/metabolism , Inflammation/complications , Inflammation/drug therapy , Inflammation/metabolism , Male , Neuroglia/drug effects , Neuroglia/metabolism , Neuroglia/pathology , Rats , Rats, Wistar , Receptors, Purinergic P2X7/metabolism , Retina/metabolism , Retina/pathology , Streptozocin , Treatment Outcome
20.
Dig Dis Sci ; 64(7): 1815-1829, 2019 07.
Article in English | MEDLINE | ID: mdl-30734238

ABSTRACT

INTRODUCTION: Our work analyzed the effects of a P2X7 receptor antagonist, Brilliant Blue G (BBG), on rat ileum myenteric plexus following ischemia and reperfusion (ISR) induced by 45 min of ileal artery occlusion with an atraumatic vascular clamp with 24 h (ISR 24-h group) or 14 d of reperfusion (ISR 14-d group). MATERIAL AND METHODS: Either BBG (50 mg/kg or 100 mg/kg, BBG50 or BBG100 groups) or saline (vehicle) was administered subcutaneously 1 h after ischemia in the ISR 24-h group or once daily for the 5 d after ischemia in the ISR 14-d group (n = 5 per group). We evaluated the neuronal density and profile area by examining the number of neutrophils in the intestinal layers, protein expression levels of the P2X7 receptor, intestinal motility and immunoreactivity for the P2X7 receptor, nitric oxide synthase, neurofilament-200, and choline acetyl transferase in myenteric neurons. RESULTS: The neuronal density and profile area were restored by BBG following ISR. The ischemic groups showed alterations in P2X7 receptor protein expression and the number of neutrophils in the intestine and decreased intestinal motility, all of which were recovered by BBG treatment. CONCLUSION: We concluded that ISR morphologically and functionally affected the intestine and that its effects were reversed by BBG treatment, suggesting the P2X7 receptor as a therapeutic target.


Subject(s)
Ileum/innervation , Mesenteric Ischemia/drug therapy , Myenteric Plexus/drug effects , Neurons/drug effects , Purinergic P2X Receptor Antagonists/pharmacology , Receptors, Purinergic P2X7/drug effects , Reperfusion Injury/prevention & control , Rosaniline Dyes/pharmacology , Animals , Cytoprotection , Disease Models, Animal , Gastrointestinal Motility/drug effects , Male , Mesenteric Ischemia/metabolism , Mesenteric Ischemia/pathology , Mesenteric Ischemia/physiopathology , Myenteric Plexus/metabolism , Myenteric Plexus/pathology , Neurons/metabolism , Neurons/pathology , Neutrophil Infiltration/drug effects , Rats, Wistar , Receptors, Purinergic P2X7/metabolism , Reperfusion Injury/metabolism , Reperfusion Injury/pathology , Reperfusion Injury/physiopathology , Signal Transduction/drug effects
SELECTION OF CITATIONS
SEARCH DETAIL