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1.
J Pharmacol Sci ; 148(1): 56-64, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34924130

RESUMEN

Acute kidney injury (AKI) is a serious complication in critically ill patients. Accumulating evidences indicated that macrophages play an important pro-inflammatory role in AKI and isoliquiritigenin (ISL) can inhibit macrophagic inflammation, but its role in AKI and the underlying mechanism are unknown. The present study aims to investigate the renoprotective effect of ISL on AKI and the role of Formyl peptide receptors 2 (FPR2) in this process. In this study, cisplatin-induced AKI model and lipopolysaccharide-induced macrophage inflammatory model were employed to perform the in vivo and in vitro experiments. The results showed that ISL strongly relieved kidney injury and inhibited renal inflammation in vivo and suppress macrophagic inflammatory response in vitro. Importantly, it was found that FPR2 was significantly upregulated compared to the control group in AKI and LPS-induced macrophage, whereas it was strongly suppressed by ISL. Interestingly, overexpression of FPR2 with transfection of pcDNA3.1-FPR2 effectively reversed the anti-inflammatory effect of ISL in macrophage, suggesting that FPR2 may be the potential target for ISL to prevent inflammation and improve kidney injury of AKI. Take together, these findings indicated that ISL improved cisplantin-induced kidney injury by inhibiting FPR2 involved macrophagic inflammation, which may provide a potential therapeutic option for AKI.


Asunto(s)
Lesión Renal Aguda/genética , Lesión Renal Aguda/prevención & control , Chalconas/farmacología , Chalconas/uso terapéutico , Cisplatino/efectos adversos , Macrófagos/metabolismo , Receptores de Formil Péptido/antagonistas & inhibidores , Receptores de Lipoxina/antagonistas & inhibidores , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/tratamiento farmacológico , Animales , Células Cultivadas , Chalconas/aislamiento & purificación , Expresión Génica/efectos de los fármacos , Glycyrrhiza/química , Inflamación , Masculino , Ratones Endogámicos C57BL , Terapia Molecular Dirigida , Fitoterapia , Receptores de Formil Péptido/genética , Receptores de Formil Péptido/metabolismo , Receptores de Formil Péptido/fisiología , Receptores de Lipoxina/genética , Receptores de Lipoxina/metabolismo , Receptores de Lipoxina/fisiología , Regulación hacia Arriba/efectos de los fármacos
2.
Biochem Pharmacol ; 190: 114596, 2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-33964283

RESUMEN

Psoriasis is a long-lasting inflammatory skin disease lacking proper cure. Dysregulated activation of neutrophils is a major pathogenic factor in psoriasis. Formyl peptide receptor 1 (FPR1) triggers neutrophil activation in response to bacteria- or mitochondria-derived N-formyl peptides, but its significance in neutrophilic psoriasis remains unknown. In this study, we discovered two derivatives of ursolic acid, 3ß-hydroxyurs-12,18-dien-28-oic acid (randialic acid B, RAB) and 3ß-hydroxyurs-12,19-dien-28-oic acid (tomentosolic acid, TA), as FPR1 inhibitors in human neutrophils with ability to suppress psoriatic symptoms in mice. Both RAB and TA, triterpenoids of traditional medicinal plant Ilex kaushue, selectively inhibited reactive oxygen species production, elastase release, and CD11b expression in human neutrophils activated by FPR1, but not non-FPR1 agonists. Importantly, RAB and TA inhibited the binding of N-formyl peptide to FPR1 in human neutrophils, neutrophil-like THP-1 cells, and hFPR1-transfected HEK293 cells, indicating FPR1 antagonism. Moreover, in assays induced by various concentrations of FPR1 agonist, both RAB and TA acted competitively for its binding to the FPR1 receptor. The FPR1-downstream signaling such as Ca2+ mobilisation and activation of Akt and MAPKs was also competitively inhibited. In addition, imiquimod-induced psoriasis-like symptoms, including epidermal hyperplasia, desquamation with scaling, neutrophil skin infiltration, and transepidermal water loss were significantly reduced by both RAB and TA. The results illustrate a possible role of human neutrophils FPR1 receptor in psoriasis-like inflammation. Accordingly, triterpenoids RAB and TA represent novel FPR1 antagonists and exhibit therapeutic potential for treating neutrophilic inflammatory skin diseases.


Asunto(s)
Neutrófilos/efectos de los fármacos , Psoriasis/prevención & control , Receptores de Formil Péptido/antagonistas & inhibidores , Triterpenos/uso terapéutico , Adulto , Animales , Línea Celular , Células Cultivadas , Femenino , Células HEK293 , Humanos , Imiquimod/toxicidad , Inflamación/inducido químicamente , Inflamación/metabolismo , Inflamación/prevención & control , Masculino , Ratones , Ratones Endogámicos BALB C , Neutrófilos/metabolismo , Psoriasis/inducido químicamente , Psoriasis/metabolismo , Receptores de Formil Péptido/metabolismo , Triterpenos/química , Triterpenos/farmacología , Adulto Joven , Ácido Ursólico
3.
Pflugers Arch ; 473(4): 683-695, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33474635

RESUMEN

The pro-resolving mechanism is a recently described endogenous process that controls inflammation. The present study evaluated components of this mechanism, including annexin 1 (ANXA1) and the formyl peptide receptor 2/ALX (FPR2/ALX) receptor, in the antihyperalgesic effect induced by electroacupuncture (EA) in an animal model of persistent peripheral inflammation. Male Swiss mice underwent intraplantar (i.pl.) injection with complete Freund's adjuvant (CFA). Mechanical hyperalgesia was assessed with von Frey monofilaments. Animals were treated with EA (2-10 Hz, ST36-SP6) or subcutaneous BML-111 injection (FPR2/ALX agonist) for 5 consecutive days. In a separate set of experiments, on the first and fifth days after CFA injection, animals received i.pl. WRW4 (FPR2/ALX antagonist) or naloxone (non-selective opioid receptor antagonist) before EA or BML-111 injection. Paw protein levels of FPR2/ALX and ANXA1 were evaluated on the second day after CFA injection by western blotting technique. EA and BML-111 reduced mechanical hyperalgesia. I.pl. naloxone or WRW4 prevented the antihyperalgesic effect induced by either EA or BML-111. EA increased ANXA1 but did not alter FPR2/ALX receptor levels in the paw. Furthermore, i.pl. pretreatment with WRW4 prevented the increase of ANXA1 levels induced by EA. This work demonstrates that the EA antihyperalgesic effect on inflammatory pain involves the ANXA1/FPR2/ALX pro-resolution pathway. This effect appears to be triggered by the activation of FPR2/ALX receptors and crosstalk communication with the opioid system.


Asunto(s)
Anexina A1/metabolismo , Electroacupuntura/métodos , Hiperalgesia/terapia , Dolor Nociceptivo/terapia , Receptores de Formil Péptido/metabolismo , Receptores Opioides/metabolismo , Animales , Adyuvante de Freund/toxicidad , Ácidos Heptanoicos/farmacología , Hiperalgesia/etiología , Hiperalgesia/metabolismo , Masculino , Ratones , Naloxona/farmacología , Antagonistas de Narcóticos/farmacología , Nocicepción/efectos de los fármacos , Dolor Nociceptivo/etiología , Dolor Nociceptivo/metabolismo , Receptores de Formil Péptido/antagonistas & inhibidores , Receptores Opioides/uso terapéutico
4.
Cell Physiol Biochem ; 51(6): 2776-2793, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30562761

RESUMEN

BACKGROUND/AIMS: Formyl peptide receptors (FPRs) recognize different endogenous and exogenous molecular stimuli and mediate neutrophil activation. Dysregulation of excessive neutrophil activation and the resulting immune responses can induce acute lung injury (ALI) in the host. Accordingly, one promising approach to the treatment of neutrophil-dominated inflammatory diseases involves therapeutic FPR1 inhibition. METHODS: We extracted a potent FPR1 antagonist from Garcinia multiflora Champ. (GMC). The inhibitory effects of GMC on superoxide anion release and elastase degranulation from activated human neutrophils were determined with spectrophotometric analysis. Reactive oxygen species (ROS) production and the FPR1 binding ability of neutrophils were assayed by flow cytometry. Signaling transduction mediated by GMC in response to chemoattractants was assessed with a calcium influx assay and western blotting. A lipopolysaccharide (LPS)-induced ALI mouse model was used to determine the therapeutic effects of GMC in vivo. RESULTS: GMC significantly reduced superoxide anion release, the reactive oxidants derived therefrom, and elastase degranulation mediated through selective, competitive FPR1 blocking in N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLF)-stimulated human neutrophils. In cell-free systems, GMC was unable to scavenge superoxide anions or suppress elastase activity. GMC produced a right shift in fMLF-activated concentration-response curves and was confirmed to be a competitive FPR1 antagonist. GMC binds to FPR1 not only in neutrophils, but also FPR1 in neutrophil-like THP-1 and hFPR1-transfected HEK293 cells. Furthermore, the mobilization of calcium and phosphorylation of mitogen-activated protein kinases and Akt, which are involved in FPR1-mediated downstream signaling, was competitively blocked by GMC. In an in vivo study, GMC significantly reduced pulmonary edema, neutrophil infiltration, and alveolar damage in LPS-induced ALI mice. CONCLUSION: Our findings demonstrate that GMC is a natural competitive FPR1 inhibitor, which makes it a possible anti-inflammatory treatment option for patients critically inflicted with FPR1-mediated neutrophilic lung damage.


Asunto(s)
Lesión Pulmonar Aguda/prevención & control , Antiinflamatorios/uso terapéutico , Garcinia/química , Activación Neutrófila/efectos de los fármacos , Extractos Vegetales/uso terapéutico , Receptores de Formil Péptido/antagonistas & inhibidores , Lesión Pulmonar Aguda/inmunología , Lesión Pulmonar Aguda/patología , Animales , Antiinflamatorios/farmacología , Línea Celular , Humanos , Ratones Endogámicos C57BL , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/patología , Extractos Vegetales/farmacología , Sustancias Protectoras/farmacología , Sustancias Protectoras/uso terapéutico , Especies Reactivas de Oxígeno/inmunología , Receptores de Formil Péptido/inmunología , Superóxidos/inmunología
5.
Sci Rep ; 7(1): 6718, 2017 07 27.
Artículo en Inglés | MEDLINE | ID: mdl-28751674

RESUMEN

Formyl peptide receptor 1 (FPR1) mediates bacterial and mitochondrial N-formyl peptides-induced neutrophil activation. Therefore, FPR1 is an important therapeutic target for drugs to treat septic or sterile inflammatory diseases. Honokiol, a major bioactive compound of Magnoliaceae plants, possesses several anti-inflammatory activities. Here, we show that honokiol exhibits an inhibitory effect on FPR1 binding in human neutrophils. Honokiol inhibited superoxide anion generation, reactive oxygen species formation, and elastase release in bacterial or mitochondrial N-formyl peptides (FPR1 agonists)-activated human neutrophils. Adhesion of FPR1-induced human neutrophils to cerebral endothelial cells was also reduced by honokiol. The receptor-binding results revealed that honokiol repressed FPR1-specific ligand N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein binding to FPR1 in human neutrophils, neutrophil-like THP-1 cells, and hFPR1-transfected HEK293 cells. However, honokiol did not inhibit FPR2-specific ligand binding to FPR2 in human neutrophils. Furthermore, honokiol inhibited FPR1 agonist-induced calcium mobilization as well as phosphorylation of p38 MAPK, ERK, and JNK in human neutrophils. In conclusion, our data demonstrate that honokiol may have therapeutic potential for treating FPR1-mediated inflammatory diseases.


Asunto(s)
Antiinflamatorios/farmacología , Compuestos de Bifenilo/farmacología , Células Endoteliales/efectos de los fármacos , Lignanos/farmacología , Neutrófilos/efectos de los fármacos , Oligopéptidos/antagonistas & inhibidores , Receptores de Formil Péptido/genética , Animales , Antiinflamatorios/aislamiento & purificación , Compuestos de Bifenilo/aislamiento & purificación , Encéfalo/citología , Encéfalo/metabolismo , Adhesión Celular/efectos de los fármacos , Técnicas de Cocultivo , Células Endoteliales/citología , Células Endoteliales/metabolismo , Regulación de la Expresión Génica , Células HEK293 , Humanos , Lignanos/aislamiento & purificación , MAP Quinasa Quinasa 4/genética , MAP Quinasa Quinasa 4/inmunología , Magnolia/química , Ratones , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/inmunología , Activación Neutrófila/efectos de los fármacos , Neutrófilos/citología , Neutrófilos/inmunología , Oligopéptidos/farmacología , Extractos Vegetales/química , Receptores de Formil Péptido/antagonistas & inhibidores , Receptores de Formil Péptido/inmunología , Receptores de Lipoxina/genética , Receptores de Lipoxina/inmunología , Células THP-1 , Proteínas Quinasas p38 Activadas por Mitógenos/genética , Proteínas Quinasas p38 Activadas por Mitógenos/inmunología
6.
BMB Rep ; 49(9): 520-5, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27502013

RESUMEN

We investigated the effects of two antimicrobial peptides (AMPs) isolated from Scolopendra subspinipes mutilans on neutrophil activity. Stimulation of mouse neutrophils with the two AMPs elicited chemotactic migration of the cells in a pertussis toxin-sensitive manner. The two AMPs also stimulated activation of ERK and Akt, which contribute to chemotactic migration of neutrophils. We found that AMP-stimulated neutrophil chemotaxis was blocked by a formyl peptide receptor (FPR) 1 antagonist (cyclosporin H); moreover the two AMPs stimulated the chemotactic migration of FPR1-expressing RBL-2H3 cells but not of vector-expressing RBL-2H3 cells. We also found that the two AMPs stimulate neutrophil migration in vivo, and that this effect is blocked in FPR1-deficient mice. Taken together, our results suggest that the two AMPs stimulate neutrophils, leading to chemotactic migration through FPR1, and the two AMPs will be useful for the study of FPR1 signaling and neutrophil activation. [BMB Reports 2016; 49(9): 520-525].


Asunto(s)
Antiinfecciosos/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Quimiotaxis de Leucocito/efectos de los fármacos , Receptores de Formil Péptido/metabolismo , Animales , Western Blotting , Línea Celular , Ciclosporina/farmacología , Alcaloides Diterpénicos , Medicamentos Herbarios Chinos/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/citología , Neutrófilos/efectos de los fármacos , Neutrófilos/metabolismo , Toxina del Pertussis/toxicidad , Fosforilación/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptores de Formil Péptido/antagonistas & inhibidores , Receptores de Formil Péptido/genética , Transducción de Señal/efectos de los fármacos
7.
Methods Mol Biol ; 910: 125-43, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22821595

RESUMEN

Virtual screening is an established technique that has successfully been deployed in the identification of novel biologically active molecules. Whether for ligand-based or for structure-based virtual screening, a chemical collection needs to be properly processed prior to in silico evaluation. Here we describe our step-by-step procedure for handling large collections of compounds prior to virtual screening.


Asunto(s)
Evaluación Preclínica de Medicamentos/métodos , Ligandos , Estructura Molecular , Receptores de Estrógenos/agonistas , Receptores de Estrógenos/antagonistas & inhibidores , Receptores de Formil Péptido/antagonistas & inhibidores , Programas Informáticos , Relación Estructura-Actividad
8.
PLoS Pathog ; 5(4): e1000362, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19343210

RESUMEN

In inflammation, pain is regulated by a balance of pro- and analgesic mediators. Analgesic mediators include opioid peptides which are secreted by neutrophils at the site of inflammation, leading to activation of opioid receptors on peripheral sensory neurons. In humans, local opioids and opioid peptides significantly downregulate postoperative as well as arthritic pain. In rats, inflammatory pain is induced by intraplantar injection of heat inactivated Mycobacterium butyricum, a component of complete Freund's adjuvant. We hypothesized that mycobacterially derived formyl peptide receptor (FPR) and/or toll like receptor (TLR) agonists could activate neutrophils, leading to opioid peptide release and inhibition of inflammatory pain. In complete Freund's adjuvant-induced inflammation, thermal and mechanical nociceptive thresholds of the paw were quantified (Hargreaves and Randall-Selitto methods, respectively). Withdrawal time to heat was decreased following systemic neutrophil depletion as well as local injection of opioid receptor antagonists or anti-opioid peptide (i.e. Met-enkephalin, beta-endorphin) antibodies indicating an increase in pain. In vitro, opioid peptide release from human and rat neutrophils was measured by radioimmunoassay. Met-enkephalin release was triggered by Mycobacterium butyricum and formyl peptides but not by TLR-2 or TLR-4 agonists. Mycobacterium butyricum induced a rise in intracellular calcium as determined by FURA loading and calcium imaging. Opioid peptide release was blocked by intracellular calcium chelation as well as phosphoinositol-3-kinase inhibition. The FPR antagonists Boc-FLFLF and cyclosporine H reduced opioid peptide release in vitro and increased inflammatory pain in vivo while TLR 2/4 did not appear to be involved. In summary, mycobacteria activate FPR on neutrophils, resulting in tonic secretion of opioid peptides from neutrophils and in a decrease in inflammatory pain. Future therapeutic strategies may aim at selective FPR agonists to boost endogenous analgesia.


Asunto(s)
Mycobacterium/inmunología , Neutrófilos/metabolismo , Nociceptores/metabolismo , Péptidos Opioides/metabolismo , Receptores de Formil Péptido/metabolismo , Análisis de Varianza , Animales , Calcio/metabolismo , Ciclosporina/metabolismo , Encefalina Metionina/metabolismo , Adyuvante de Freund/metabolismo , Humanos , Masculino , Monocitos/metabolismo , Antagonistas de Narcóticos , Neurotransmisores/metabolismo , Dolor/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Ratas , Ratas Wistar , Receptores de Formil Péptido/agonistas , Receptores de Formil Péptido/antagonistas & inhibidores , Receptores Toll-Like/agonistas
9.
Chem Biol Interact ; 179(2-3): 419-29, 2009 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-19167369

RESUMEN

Gliomas are the most common and lethal tumor type in the brain. The present study investigated the effect of oligomer procyanidins (F2) (F2, degree of polymerization 2-15), a natural fraction isolated from grape seeds on the biological behavior of glioblastoma cells. We found that F2 significantly inhibited the glioblastoma growth, with little cytotoxicity on normal cells, induced G2/M arrest and decreased mitochondrial membrane potential in U-87 cells. It also induced a non-apoptotic cell death phenotype resembling paraptosis in U-87 cells. In addition, it was found for the first time that F2 in non-cytotoxic concentrations selectively inhibited U-87 cell chemotaxis mediated by a G-protein coupled receptor formyl peptide receptor FPR, which is implicated in tumor cell invasion and metastasis. Further experiments indicated that F2 inhibited fMLF-induced U-87 cell calcium mobilization and MAP kinases ERK1/2 phosphorylation. Moreover, F2 attenuated the glioblastoma FPR expression, a new molecular target for glioma therapeutics, which has been shown to play important roles in glioma cells chemotaxis, proliferation and angiogenesis in addition to its promotion to tumor progression, but did not affect FPR mRNA expression in U-87 cells. Taken together, our results suggest that F2 may be a promising candidate for the development of novel anti-tumor therapeutics.


Asunto(s)
Glioblastoma/metabolismo , Glioblastoma/patología , Extractos Vegetales/farmacología , Proantocianidinas/farmacología , Receptores de Formil Péptido/antagonistas & inhibidores , Semillas/química , Vitis/química , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Membranas Mitocondriales/efectos de los fármacos , Membranas Mitocondriales/metabolismo , Estructura Molecular , Proantocianidinas/química , Proantocianidinas/aislamiento & purificación , Receptores de Formil Péptido/metabolismo , Células Tumorales Cultivadas
10.
Mol Pharmacol ; 68(5): 1301-10, 2005 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-16118363

RESUMEN

The formylpeptide receptor (FPR) family of G-protein-coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. We developed a FPR homology model and pharmacophore (based on the bovine rhodopsin crystal structure and known FPR ligands, respectively) for in silico screening of approximately 480,000 drug-like small molecules. A subset of 4324 compounds that matched the pharmacophore was then physically screened with the HyperCyt flow cytometry platform in high-throughput, no-wash assays that directly measure human FPR binding, with samples (each approximately 2500 cells in 2 microl) analyzed at 40/min. From 52 confirmed hits (1.2% hit rate), we identified 30 potential lead compounds (inhibition constant, Ki= 1-32 microM) representing nine distinct chemical families. Four compounds in one family were weak partial agonists. All others were antagonists. This virtual screening approach improved the physical screening hit rate by 12-fold (versus 0.1% hit-rate in a random compound collection), providing an efficient process for identifying small molecule antagonists.


Asunto(s)
Evaluación Preclínica de Medicamentos , Citometría de Flujo/métodos , Receptores de Formil Péptido/antagonistas & inhibidores , Secuencia de Aminoácidos , Animales , Bovinos , Modelos Moleculares , Datos de Secuencia Molecular , Receptores de Formil Péptido/agonistas , Receptores de Formil Péptido/química
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