RESUMEN

The natural compound ravenelin was isolated from the biomass extracts of Exserohilum rostratum fungus, and its antimicrobial, antiplasmodial, and trypanocidal activities were evaluated. Ravenelin was isolated by column chromatography and HPLC and identified by NMR and MS. The susceptibility of Gram-positive and Gram-negative bacteria strains to ravenelin was determined by microbroth dilution assay. Cytotoxicity was evaluated in hepatocarcinoma cells (HepG2) and BALB/c peritoneal macrophages by using MTT. SYBR Green I-based assay was used in the asexual stages of Plasmodium falciparum. Trypanocidal activity was tested against the epimastigote and intracellular amastigote forms of Trypanosoma cruzi. Ravenelin was active against Gram-positive bacteria strains, with emphasis on Bacillus subtilis (MIC value of 7.5 µM). Ravenelin's antiparasitic activities were assessed against both the epimastigote (IC50 value of 5 ± 1 µM) and the intracellular amastigote forms of T. cruzi (IC50 value of 9 ± 2 µM), as well as against P. falciparum (IC50 value of 3.4 ± 0.4 µM). Ravenelin showed low cytotoxic effects on both HepG2 (CC50 > 50 µM) and peritoneal macrophage (CC50 = 185 ± 1 µM) cells with attractive selectivity for the parasites (SI values > 15). These findings indicate that ravenelin is a natural compound with both antibacterial and antiparasitic activities, and considerable selectivity indexes. Therefore, ravenelin is an attractive candidate for hit-to-lead development.

Asunto(s)

Antibacterianos/farmacología , Antiprotozoarios/farmacología , Ascomicetos/química , Macrófagos Peritoneales/citología , Xantonas/farmacología , Animales , Antibacterianos/química , Antiprotozoarios/química , Productos Biológicos/química , Productos Biológicos/farmacología , Biomasa , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Cromatografía Líquida de Alta Presión , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Células Hep G2 , Humanos , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/parasitología , Espectroscopía de Resonancia Magnética , Ratones , Ratones Endogámicos BALB C , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Plasmodium falciparum/efectos de los fármacos , Trypanosoma cruzi/efectos de los fármacos , Xantonas/química

RESUMEN

Trypanosoma cruzi is the causative agent of Chagas disease, which is endemic in Latin America and around the world through mother to child transmission. The heart is the organ most frequently affected in the chronic stage of the human infection and depends on mitochondria for the required energy for its activity. Cyclophilins are involved in protein folding and the mitochondrial isoform, Cyclophilin D (CyPD), has a crucial role in the opening of the mitochondrial permeability transition pore. In the present study, we infected CyPD deficient mice, with ablation of the Ppif gene, with T. cruzi parasites and the course of the infection was analyzed. Parasite load, quantified by PCR, was significantly lower in skeletal and cardiac tissues of Ppif-/- mice compared to wild type mice. In vitro cultured cardiomyocytes and macrophages from mice lacking CyPD exhibited lower percentage of infected cells and number of intracellular parasites than those observed for wild type mice. Although histopathological analysis of heart and mRNA of heart cytokines showed differences between T. cruzi-infected mice compared to the uninfected animals, no significant differences were found mice due to the ablation of the Ppif gene. Our results suggest that cells deficient for mitochondrial CyPD, inhibited for the mitochondrial membrane potential collapse, reduces the severity of parasite aggression and spread of cellular infection.

Asunto(s)

Enfermedad de Chagas/parasitología , Peptidil-Prolil Isomerasa F/deficiencia , Trypanosoma cruzi/fisiología , Animales , Citocinas/análisis , Citocinas/genética , ADN Protozoario/aislamiento & purificación , Corazón/parasitología , Hígado/patología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/parasitología , Ratones , Ratones Endogámicos C57BL , Músculo Esquelético/parasitología , Músculo Esquelético/patología , Miocardio/patología , Miocitos Cardíacos/citología , Miocitos Cardíacos/parasitología , Carga de Parásitos , ARN Mensajero/análisis , ARN Protozoario/análisis , ARN Protozoario/aislamiento & purificación , Bazo/patología , Trypanosoma cruzi/genética

RESUMEN

Leishmaniasis is a disease impacting public health worldwide due to its high incidence, morbidity and mortality. Available treatments are costly, lengthy and toxic, not to mention the problem of parasite resistance. The development of alternative treatments is warranted and natural products demonstrate promising activity. This study investigated the activity of Connarus suberosus extracts and compounds against Leishmania species. Several C. suberosus extracts were tested against L. amazonensis promastigotes. Active and inactive extracts were analyzed by UHPLC-MS and data evaluated using a metabolomics platform, revealing an unknown neoflavonoid (connarin, 3), isolated together with the pterocarpans: hemileiocarpin (1) and leiocarpin (2). The aforementioned compounds (1-3), together with the benzoquinones: rapanone (4), embelin (5) and suberonone (6) previously isolated by our group from the same species, were tested against: (i) L. amazonensis and L. infantum promastigotes, and (ii) L. amazonensis intracellular amastigotes, with the most active compound (3) also tested against L. infantum amastigotes. Cytotoxicity against murine peritoneal macrophages was also investigated. Compounds 2 and 3 presented an IC50 33.8 µM and 11.4 µM for L. amazonensis promastigotes; and 44.3 µM and 13.3 µM for L. infantum promastigotes, respectively. For L. amazonensis amastigotes, the IC50 of 2 was 20.4 µM with a selectivity index (SI) of 5.7, while the IC50 of 3 was 2.9 µM with an SI of 6.3. For L. infantum amastigotes, the IC50 of 3 was 7.7 µM. Compounds 2 and 3 presented activity comparable with the miltefosine positive control, with compound 3 found to be 2-4 times more active than the positive control, depending on the Leishmania species and form. The extracts and isolated compounds showed moderate toxicity against macrophages. Compounds 2 and 3 altered the mitochondrial membrane potential (ΔΨm) and neutral lipid body accumulation, while 2 also impacted plasma membrane permeabilization, culminating in cellular disorder and parasite death. Transmission electron microscopy of L. amazonensis promastigotes treated with compound 3 confirmed the presence of lipid bodies. Leiocarpin (2) and connarin (3) demonstrated antileishmanial activity. This study provides knowledge of natural products with antileishmanial activity, paving the way for prototype development to fight this neglected tropical disease.

Asunto(s)

Connaraceae/química , Flavonoides/farmacología , Metabolómica/métodos , Extractos Vegetales/farmacología , Animales , Antiprotozoarios/química , Antiprotozoarios/aislamiento & purificación , Antiprotozoarios/farmacología , Supervivencia Celular , Cromatografía Líquida de Alta Presión , Flavonoides/química , Flavonoides/aislamiento & purificación , Leishmania mexicana/efectos de los fármacos , Leishmania mexicana/crecimiento & desarrollo , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/parasitología , Espectrometría de Masas , Ratones , Ratones Endogámicos BALB C , Estructura Molecular , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación

RESUMEN

There are different varieties of mushrooms not yet studied spread all over the planet. The objective of this study was to evaluate biochemical properties and effects on mitochondrial respiration of eight Basidiomycete mushrooms: Flaviporus venustus EF30, Hydnopolyporus fimbriatus EF41 and EF44, Inonotus splitgerberi EF46, Oudemansiella canarii EF72, Perenniporia sp. EF79, Phellinus linteus EF81, and Pleurotus albidus EF84. Total phenols, ABTS, TEAC, FRAP, and ORAC were measured in order to determine the antioxidant capacity. Antimicrobial potential was studied by disc-diffusion and microdilution method. Cytotoxicity was determined in murine peritoneal macrophages. The bioenergetic aspects were evaluated by the uncoupling of the oxidative phosphorylation in mitochondrias. The H. fimbriatus mushroom was the one that presented the most significant results for the antioxidant assays. Three mushrooms presented antimicrobial activity, indicating a potential for formulation of drugs. The results suggest that I. spligerberi has an uncoupling activity, even at the lowest concentration tested, dissipating the mitochondrial electrochemical gradient. On the other hand, P. albidus has effect only on succinate-oxidase activity without influencing mitochondrial respiratory efficiency. Therefore, both interfere negatively in mitochondrial respiration. In relation with the cytotoxicity in peritoneal macrophages, O. canarii and F. venustus were cytotoxic in this type of cells.

Asunto(s)

Basidiomycota/química , Mitocondrias/efectos de los fármacos , Fenoles/química , Animales , Antibacterianos/química , Antibacterianos/farmacología , Antioxidantes/química , Antioxidantes/metabolismo , Basidiomycota/metabolismo , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Mitocondrias/metabolismo , Oxidación-Reducción , Fosforilación Oxidativa/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Fenoles/aislamiento & purificación , Fenoles/farmacología , Staphylococcus aureus/efectos de los fármacos , Ácido Succínico/química

RESUMEN

Photodynamic therapy has emerged as an alternative treatment for cutaneous leishmaniasis, and compounds with photocatalytic behavior are promising candidates to develop new therapeutic strategies for the treatment of this parasitic disease. Titanium dioxide TiO2 is a semiconductor ceramic material that shows excellent photocatalytic and antimicrobial activity under Ultraviolet irradiation. Due to the harmful effects of UV radiation, many efforts have been made in order to enhance both photocatalytic and antimicrobial properties of TiO2 in the visible region of the spectrum by doping or through modifications in the route of synthesis. Herein, Fe-, Zn-, or Pt- doped TiO2 nanostructures were synthesized by solution-combustion route. The obtained compounds presented aggregates of 100 nm, formed by particles smaller than 20 nm. Doping compounds shift the absorption spectrum towards the visible region, allowing production of reactive oxygen species in the presence of oxygen and molecular water when the system is irradiated in the visible spectrum. The Pt (EC50 = 18.2 ±â€¯0.8 µg/mL) and Zn (EC50 = 16.4 ±â€¯0.3 µg/mL) -doped TiO2 presented the higher antileishmanial activities under visible irradiation and their application as photosensitizers in photodynamic therapy (PDT) strategies for the treatment of cutaneous leishmaniasis should be considered.

Asunto(s)

Leishmania/efectos de los fármacos , Nanopartículas del Metal/toxicidad , Fármacos Fotosensibilizantes/farmacología , Titanio/química , Rayos Ultravioleta , Animales , Antracenos , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Daño del ADN/efectos de los fármacos , Daño del ADN/efectos de la radiación , Hierro/química , Leishmania/metabolismo , Leishmania/efectos de la radiación , Leishmaniasis/tratamiento farmacológico , Leishmaniasis/parasitología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Macrófagos Peritoneales/efectos de la radiación , Nanopartículas del Metal/química , Nanopartículas del Metal/uso terapéutico , Ratones , Microscopía Electrónica de Transmisión , Perileno/análogos & derivados , Perileno/química , Perileno/farmacología , Perileno/uso terapéutico , Fotoquimioterapia , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/uso terapéutico , Especies Reactivas de Oxígeno/metabolismo , Espectrometría de Fluorescencia , Difracción de Rayos X , Zinc/química

RESUMEN

4-(Nitrophenyl)hydrazone derivatives of N-acylhydrazone were synthesized and screened for suppress lymphocyte proliferation and nitrite inhibition in macrophages. Compared to an unsubstituted N-acylhydrazone, active compounds were identified within initial series when hydroxyl, chloride and nitro substituents were employed. Structure-activity relationship was further developed by varying the position of these substituents as well as attaching structurally-related substituents. Changing substituent position revealed a more promising compound series of anti-inflammatory agents. In contrast, an N-methyl group appended to the 4-(nitrophenyl)hydrazone moiety reduced activity. Anti-inflammatory activity of compounds is achieved by modulating IL-1ß secretion and prostaglandin E2 synthesis in macrophages and by inhibiting calcineurin phosphatase activity in lymphocytes. Compound SintMed65 was advanced into an acute model of peritonitis in mice, where it inhibited the neutrophil infiltration after being orally administered. In summary, we demonstrated in great details the structural requirements and the underlying mechanism for anti-inflammatory activity of a new family of hydrazone-N-acylhydrazone, which may represent a valuable medicinal chemistry direction for the anti-inflammatory drug development in general.

Asunto(s)

Antiinflamatorios/síntesis química , Diseño de Fármacos , Hidrazonas/química , Factores Inmunológicos/síntesis química , Animales , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Supervivencia Celular/efectos de los fármacos , Cristalografía por Rayos X , Dinoprostona/metabolismo , Modelos Animales de Enfermedad , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Hidrazonas/farmacología , Hidrazonas/uso terapéutico , Factores Inmunológicos/farmacología , Factores Inmunológicos/uso terapéutico , Interleucina-1beta/metabolismo , Lipopolisacáridos/toxicidad , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Conformación Molecular , Óxido Nítrico/metabolismo , Peritonitis/tratamiento farmacológico , Peritonitis/patología , Relación Estructura-Actividad

RESUMEN

Inflammation is a major local feature of envenomation by bothropic snakes being characterized by a prominent local edema, pain, and extensive swelling. There are reports demonstrating that whole Bothrops snake venoms and toxins isolated from them are able to activate macrophages functions, such as phagocytosis, production of reactive oxygen, cytokines and eicosanoids, however, little is known about the effects of Bothrops alternatus (B.a.) venom on macrophages. In this work, we evaluated the proinflammatory effects of B.a. venom with in vivo and in vitro experiments using the Raw 264.7 cell line and mouse peritoneal macrophages. We detected that B.a. venom augments cell permeability (2-fold), and cellular extravasation (mainly neutrophils), increase proinflammatory cytokines IL1 (∼300-fold), IL12 (∼200-fold), and TNFα (∼80-fold) liberation and induce the expression of enzymes related to lipid signaling, such as cPLA2α and COX-2. Additionally, using lipidomic techniques we detected that this venom produces a release of arachidonic acid (∼10 nMol/mg. Protein) and other fatty acids (16:0 and 18:1 n-9c). Although much of these findings were described in inflammatory processes induced by other bothropic venoms, here we demonstrate that B.a. venom also stimulates pro-inflammatory pathways involving lipid mediators of cell signaling. In this sense, lipidomics analysis of macrophages stimulated with B.a. venom evidenced that the main free fatty acids are implicated in the inflammatory response, and also demonstrated that this venom, is able to activate lipid metabolism even with a low content of PLA2.

Asunto(s)

Bothrops/metabolismo , Macrófagos Peritoneales/efectos de los fármacos , Venenos de Serpiente/toxicidad , Animales , Ácido Araquidónico/análisis , Ácido Araquidónico/metabolismo , Permeabilidad de la Membrana Celular/efectos de los fármacos , Células Cultivadas , Ciclooxigenasa 2/metabolismo , Citocinas/metabolismo , Edema/etiología , Ácidos Grasos/análisis , Ácidos Grasos/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Fosfolipasas A2 Grupo IV/metabolismo , Interleucina-1/metabolismo , Interleucina-12/metabolismo , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/metabolismo , Masculino , Ratones , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/metabolismo , Células RAW 264.7 , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Tissue macrophages are a heterogeneous cell population residing in all body tissues that contribute to the maintenance of homeostasis and trigger immune activation in response to injurious stimuli. This heterogeneity may be associated with tissue-specific functions; however, the presence of distinct macrophage populations within the same microenvironment indicates that macrophage heterogeneity may also be influenced outside of tissue specialization. The F4/80 molecule was established as a unique marker of murine macrophages when a monoclonal antibody was found to recognize an antigen exclusively expressed by these cells. However, recent research has shown that F4/80 is expressed by other immune cells and is not equivalently expressed across tissue-specific macrophage lineages, including those residing in the same microenvironment, such as the peritoneum and spleen. In this context, two murine macrophage subtypes with distinct F4/80 expression patterns were recently found to coexist in the peritoneum, termed large peritoneal macrophages (LPMs) and small peritoneal macrophages (SPMs). However, the presence of phenotypic and functional heterogeneous macrophage subpopulations in the spleen was already known. Thus, although F4/80 surface expression continues to be the best method to identify tissue macrophages, additional molecules must also be examined to distinguish these cells from other immune cells.

Asunto(s)

Antígenos de Diferenciación/biosíntesis , Biomarcadores/metabolismo , Macrófagos Peritoneales/citología , Bazo/citología , Animales , Humanos , Macrófagos/citología , Macrófagos/metabolismo , Macrófagos Peritoneales/metabolismo , Ratones , Peritoneo/citología

RESUMEN

In vitro effects of acetylated pectin (OP) isolated from cacao pod husks (Theobroma cacao L.), its partially deacetylated and de-esterified form (MOP), and a commercial homogalacturonan (PG) were investigated on murine peritoneal macrophages. MOP stood out among the studied pectins. After 48h of incubation, compared with the control group, it was able to promote significant macrophage morphological differentiation from resident to activated stage and also stimulated nitric oxide production, which reached a level of 85% of that of LPS stimulus. In the presence of the highest tested concentration of MOP (200µg·mL-1), the levels of the cytokines TNF-α (6h) and IL-12 and IL-10 (48h) increased substantially in relation to untreated cells. Our results show that the partial deacetylation and de-esterification of pectin extracted from cacao pod husks (T. cacao L.) produced a polymer with greater ability than its native form to activate macrophages to a cytotoxic phenotype. Like this, they provide the possibility of a therapeutic application to MOP, which could lead to a decreased susceptibility to microbial infection besides antitumor activity. Additionally, the present results also corroborate with the proposition of that the chemical modifications of the biopolymers can result in an improved molecule with new possibilities of application.

Asunto(s)

Cacao/química , Activación de Macrófagos/efectos de los fármacos , Macrófagos Peritoneales/efectos de los fármacos , Pectinas/farmacología , Acetilación , Animales , Femenino , Expresión Génica , Inflamación , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-12/genética , Interleucina-12/inmunología , Lipopolisacáridos/farmacología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/inmunología , Ratones , Óxido Nítrico/agonistas , Óxido Nítrico/biosíntesis , Pectinas/química , Pectinas/aislamiento & purificación , Cultivo Primario de Células , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunología

RESUMEN

AIMS: The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that mediates the toxicity of environmental pollutants. It is also implicated in the regulation of the immune system. Ahr-null macrophages overproduce several proinflammatory cytokines following LPS-mediated stimulation, suggesting that AHR affects the balance between the inflammatory M1 and anti-inflammatory M2 phenotypes. Therefore, the present study aimed to examine whether the loss of AHR modifies macrophage polarization. MATERIALS AND METHODS: Peritoneal macrophages from wild-type and Ahr-null mice were differentiated into M1 or M2 phenotype by treatment with LPS/IFNγ or IL-4, and several M1 and M2 markers were determined by qPCR and ELISA assays. Macrophage phagocytic capacity was determined through phagocytosis of yeast and Leishmania major infection assays. Nitric oxide (NO) and urea production, and arginase activity were also determined. KEY FINDINGS: When macrophages were polarized to the M1 phenotype, Ahr-null cells presented a mixed response; higher levels of IL-1ß, IL-6, IL-12, and TNFα were observed after IFNγ- and LPS-mediated activation. However, Ahr-null cells also exhibited decreased NO production and phagocytic capacity. When macrophage was polarized to the M2 phenotype, Ahr-null cells exhibited lower levels of Fizz1, Ym1, and IL-10. In contrast, arginase activity was increased when compared to wild-type macrophages. In addition, macrophages from Ahr-null mice were more susceptible to L. major infection. SIGNIFICANCE: Disruption of the Ahr gene alters macrophage polarization when compared to WT macrophage. These changes may affect the development and resolution of several diseases such as bacterial or parasitic infections.

Asunto(s)

Arginina/biosíntesis , Polaridad Celular/fisiología , Macrófagos Peritoneales/citología , Óxido Nítrico/biosíntesis , Receptores de Hidrocarburo de Aril/fisiología , Animales , Células Cultivadas , Macrófagos Peritoneales/inmunología , Masculino , Ratones , Ratones Noqueados , Fagocitosis , Receptores de Hidrocarburo de Aril/genética

RESUMEN

Macrophages respond to endogenous and non-self stimuli acquiring the M1 or M2 phenotypes, corresponding to classical or alternative activation, respectively. The role of B-1 cells in the regulation of macrophage polarization through the secretion of interleukin (IL)-10 has been demonstrated. However, the influence of B-1 cells on macrophage phenotype induction by an immunogen that suppress their ability to secrete IL-10 has not been explored. Here, we studied the peritoneal macrophage pattern induced by liposomes comprised of dipalmitoylphosphatidylcholine (DPPC) and cholesterol (Chol) carrying ovalbumin (OVA) (Lp DPPC/OVA), and the involvement of B-1 cells in macrophage polarization. Peritoneal cells from BALB/c, B-1 cells-deficient BALB/xid and C57BL/6 mice immunized with Lp DPPC/OVA and OVA in soluble form (PBS/OVA) were analyzed and stimulated or not in vitro with lipopolysaccharide (LPS). Peritoneal macrophages from BALB/c and C57BL/6 mice immunized with Lp DPPC/OVA showed an M2-like phenotype as evidenced by their high arginase activity without LPS stimulation. Upon stimulation, these macrophages were reprogrammable toward the M1 phenotype with the upregulation of nitric oxide (NO) and a decrease in IL-10 secretion. In addition, high IFN-γ levels were detected in the culture supernatant of peritoneal cells from BALB/c and C57BL/6 mice immunized with Lp DPPC/OVA. Nevertheless, still high levels of arginase activity and undetectable levels of IL-12 were found, indicating that the switch to a classical activation state was not complete. In the peritoneal cells from liposomes-immunized BALB/xid mice, levels of arginase activity, NO, and IL-6 were below those from wild type animals, but the last two products were restored upon adoptive transfer of B-1 cells, together with an increase in IFN-γ secretion. Summarizing, we have demonstrated that Lp DPPC/OVA induce an M2-like pattern in peritoneal macrophages reprogrammable to M1 phenotype after LPS stimulation, with the involvement of B-1 cells.

Asunto(s)

Linfocitos B/inmunología , Colesterol/farmacología , Liposomas/farmacología , Macrófagos Peritoneales/inmunología , Ovalbúmina/farmacología , 1,2-Dipalmitoilfosfatidilcolina/farmacología , Traslado Adoptivo , Animales , Arginasa/biosíntesis , Linfocitos B/trasplante , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Portadores de Fármacos/farmacología , Interferón gamma/biosíntesis , Interleucina-10/metabolismo , Interleucina-12/biosíntesis , Interleucina-6/biosíntesis , Lipopolisacáridos/inmunología , Activación de Macrófagos/inmunología , Macrófagos Peritoneales/citología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Óxido Nítrico/biosíntesis , Fenotipo , Fosfatidilcolinas/farmacología

RESUMEN

Continuing our screening program for novel anti-parasite compounds, we synthesized seven 1,4-naphthoquinones coupled to 1,2,3-triazoles, five nor-ß-lapachone-based 1,2,3-triazoles and ten α-lapachone-based 1,2,3-triazoles. These and other naphthoquinonoid compounds were evaluated for their activity against promastigote forms of antimony-sensitive and -resistant strains of Leishmania infantum (syn. Leishmania chagasi) and Leishmania amazonensis. The toxicity of these compounds to mammalian cells was also examined. The substances were more potent than an antimonial drug, with IC50 values ranging from 1.0 to 50.7 µM. Nor-α-lapachone derivatives showed the highest antileishmanial activity, with selectivity indices in the range of 10-15. These compounds emerged as important leads for further investigation as antileishmanial agents. Additionally, one of these compounds exhibited cross-resistance in Sb-resistant Leishmania and could provide a molecular tool for investigating the multidrug resistance mechanisms in Leishmania parasites.

Asunto(s)

Antiprotozoarios/síntesis química , Reacción de Cicloadición/métodos , Naftoquinonas/síntesis química , Triazoles/síntesis química , Alquinos/química , Animales , Antimonio/farmacología , Antiprotozoarios/química , Antiprotozoarios/farmacología , Azidas/química , Catálisis , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Cobre/química , Resistencia a Medicamentos/efectos de los fármacos , Leishmania/efectos de los fármacos , Leishmania infantum/efectos de los fármacos , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/efectos de los fármacos , Ratones , Naftoquinonas/química , Naftoquinonas/farmacología , Pruebas de Sensibilidad Parasitaria , Especificidad de la Especie , Triazoles/química , Triazoles/farmacología

RESUMEN

Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma; it accounts for more than 280,000 deaths annually. In this work we investigated the effect of the alkaloidic extract obtained by acid-base extraction of the dried fruits of Solanum lycocarpum on schistosomiasis. We used this extract at concentrations of 10, 20, and 40 mg/kg to treat mice infected with Schistosoma mansoni in different phases of the parasite cycle, and we compared its effect with that of the positive control praziquantel (60 mg/kg). We evaluated the results on the basis of the number of macrophages, eggs, and granulomas; we also assessed nitric oxide (NO) and interferon-gamma (IFN-γ) production. Animals treated with a daily dose of 10 or 20 mg/kg alkaloidic extract between the 37th and 41st day of infection showed increased number of macrophages, elevated NO and IFN-γ concentrations, and reduced number of eggs and granulomas in the liver. The alkaloidic extract of S. lycocarpum fruits displayed an immunomodulatory effect on mice infected with S. mansoni, so its potential to treat schistosomiasis deserves further studies.

Asunto(s)

Frutas/química , Factores Inmunológicos/farmacología , Schistosoma mansoni/efectos de los fármacos , Esquistosomiasis mansoni/tratamiento farmacológico , Alcaloides Solanáceos/farmacología , Solanum/química , Animales , Antihelmínticos/farmacología , Antihelmínticos/uso terapéutico , Recuento de Células , Femenino , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/uso terapéutico , Interferón gamma/sangre , Interferón gamma/metabolismo , Hígado/parasitología , Hígado/patología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Ratones , Óxido Nítrico/metabolismo , Recuento de Huevos de Parásitos , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Praziquantel/farmacología , Praziquantel/uso terapéutico , Schistosoma mansoni/inmunología , Esquistosomiasis mansoni/inmunología , Alcaloides Solanáceos/aislamiento & purificación , Alcaloides Solanáceos/uso terapéutico

RESUMEN

The objective of this work was to determine the hematological parameters and the phagocytic capacity of peritoneal macrophages of fat snook related to sex, stage of gonadal maturation and seasonal cycle. Blood was collected from 135 animals (78 females and 57 males) and used for determinations of: erythrocyte number, hematocrit, hemoglobin, erythrocyte indices mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), total and differential leukocyte counts, and thrombocyte count. The phagocytic capacity and phagocytic index were determined after Saccharomyces cerevisiae inoculation in the peritoneal cavity of the animals. The hematological results according to sex showed that the erythrocyte, total leukocyte and thrombocyte counts were statistically higher in males than females, with the latter showing a higher MCV. Concerning to erythrocyte count, hematocrit and hemoglobin concentration analyzed separately by sex and stage of gonadal maturation, males were found to have significantly elevated values in the mature stage and decreased levels in the resting stage. The results of the erythrocyte and leukocyte series, thrombocytes and phagocytic activity related to seasonal cycle showed significant differences in both sexes, where hematocrit and hemoglobin concentration were lower in winter and higher in the other seasons, mean corpuscular volume was higher in the summer and lower in the winter and fall, total leukocytes and thrombocytes lower in the spring and higher in the fall, lymphocytes low in the winter and summer and high in the spring and phagocytic capacity and phagocytic index high in the summer and low in the winter and fall. The results showed that the hematological values in males are statistically higher than those in females, the erythrocyte values in males increase with the progression of gonadal maturation and that winter is the season of the year least favorable for hematological and phagocytic responses for survival of fat snook kept in captivity. The parameters studied could be utilized in the evaluation of the health status of this species in captivity.

Asunto(s)

Hematócrito , Macrófagos Peritoneales/inmunología , Perciformes/sangre , Perciformes/inmunología , Fagocitosis , Factores de Edad , Animales , Acuicultura , Análisis Químico de la Sangre , Recuento de Eritrocitos , Índices de Eritrocitos , Femenino , Recuento de Leucocitos , Macrófagos Peritoneales/citología , Masculino , Estaciones del Año , Factores Sexuales

RESUMEN

The inflammatory response is driven by signals that recruit and elicit immune cells to areas of tissue damage or infection. The concept of a mononuclear phagocyte system postulates that monocytes circulating in the bloodstream are recruited to inflamed tissues where they give rise to macrophages. A recent publication demonstrated that the large increase in the macrophages observed during infection was the result of the multiplication of these cells rather than the recruitment of blood monocytes. We demonstrated previously that B-1 cells undergo differentiation to acquire a mononuclear phagocyte phenotype in vitro (B-1CDP), and we propose that B-1 cells could be an alternative origin for peritoneal macrophages. A number of recent studies that describe the phagocytic and microbicidal activity of B-1 cells in vitro and in vivo support this hypothesis. Based on these findings, we further investigated the differentiation of B-1 cells into phagocytes in vivo in response to LPS-induced inflammation. Therefore, we investigated the role of B-1 cells in the composition of the peritoneal macrophage population after LPS stimulation using osteopetrotic mice, BALB/Xid mice and the depletion of monocytes/macrophages by clodronate treatment. We show that peritoneal macrophages appear in op/op((-/-)) mice after LPS stimulation and exhibit the same Ig gene rearrangement (VH11) that is often found in B-1 cells. These results strongly suggest that op/op((-/-)) peritoneal "macrophages" are B-1CDP. Similarly, the LPS-induced increase in the macrophage population was observed even following monocyte/macrophage depletion by clodronate. After monocyte/macrophage depletion by clodronate, LPS-elicited macrophages were observed in BALB/Xid mice only following the transfer of B-1 cells. Based on these data, we confirmed that B-1 cell differentiation into phagocytes also occurs in vivo. In conclusion, the results strongly suggest that B-1 cell derived phagocytes are a component of the LPS-elicited peritoneal macrophage population.

Asunto(s)

Subgrupos de Linfocitos B/inmunología , Lipopolisacáridos/inmunología , Macrófagos Peritoneales/inmunología , Fagocitos/inmunología , Animales , Subgrupos de Linfocitos B/citología , Diferenciación Celular , Células Cultivadas , Genes de Inmunoglobulinas , Inflamación/inmunología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Fagocitos/citología , Fagocitos/metabolismo

RESUMEN

5-Lipoxygenase-derived products have been implicated in both the inhibition and promotion of chronic infection. Here, we sought to investigate the roles of endogenous 5-lipoxygenase products and exogenous leukotrienes during Histoplasma capsulatum infection in vivo and in vitro. 5-LO deficiency led to increased lung CFU, decreased nitric oxide production and a deficient primary immune response during active fungal infection. Moreover, H. capsulatum-infected 5-LO(-/-) mice showed an intense influx of neutrophils and an impaired ability to generate and recruit effector T cells to the lung. The fungal susceptibility of 5-LO(-/-) mice correlated with a lower rate of macrophage ingestion of IgG-H. capsulatum relative to WT macrophages. Conversely, exogenous LTB4 and LTC4 restored macrophage phagocytosis in 5-LO deficient mice. Our results demonstrate that leukotrienes are required to control chronic fungal infection by amplifying both the innate and adaptive immune response during histoplasmosis.

Asunto(s)

Araquidonato 5-Lipooxigenasa/fisiología , Histoplasma/inmunología , Histoplasmosis/inmunología , Pulmón/inmunología , Macrófagos Peritoneales/inmunología , Linfocitos T/inmunología , Animales , Proliferación Celular , Células Cultivadas , Citotoxicidad Inmunológica , Citometría de Flujo , Histoplasmosis/microbiología , Histoplasmosis/mortalidad , Inmunidad Humoral , Inmunidad Innata , Leucotrieno B4/metabolismo , Leucotrieno C4/metabolismo , Pulmón/microbiología , Activación de Linfocitos , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/microbiología , Ratones , Ratones Noqueados , Óxido Nítrico/metabolismo , Fagocitosis , Tasa de Supervivencia , Linfocitos T/microbiología

RESUMEN

It has been previously shown that besides its classical role in blood pressure control the renin-angiotensin system, mainly by action of angiotensin II on the AT(1) receptor, exerts pro-inflammatory effects such as by inducing the production of cytokines. More recently, alternative pathways to this system were described, such as binding of angiotensin-(1-7) to receptor Mas, which was shown to counteract some of the effects evoked by activation of the angiotensin II-AT(1) receptor axis. Here, by means of different molecular approaches we investigated the role of angiotensin-(1-7) in modulating inflammatory responses triggered in mouse peritoneal macrophages. Our results show that receptor Mas transcripts were up-regulated by eightfold in LPS-induced macrophages. Interestingly, macrophage stimulation with angiotensin-(1-7), following to LPS exposure, evoked an attenuation in expression of TNF-α and IL-6 pro-inflammatory cytokines; where this event was abolished when the receptor Mas selective antagonist A779 was also included. We then used heterologous expression of the receptor Mas in HEK293T cells to search for the molecular mechanisms underlying the angiotensin-(1-7)-mediated anti-inflammatory responses by a kinase array; what suggested the involvement of the Src kinase family. In LPS-induced macrophages, this finding was corroborated using the PP2 compound, a specific Src kinase inhibitor; and also by Western blotting when we observed that Ang-(1-7) attenuated the phosphorylation levels of Lyn, a member of the Src kinase family. Our findings bring evidence for an anti-inflammatory role for angiotensin-(1-7) at the cellular level, as well as show that its probable mechanism of action includes the modulation of Src kinases activities.

Asunto(s)

Angiotensina I/farmacología , Lipopolisacáridos/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/inmunología , Fragmentos de Péptidos/farmacología , Angiotensina I/inmunología , Animales , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Células HEK293 , Humanos , Lipopolisacáridos/inmunología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Fragmentos de Péptidos/inmunología , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Familia-src Quinasas/genética , Familia-src Quinasas/metabolismo

RESUMEN

BACKGROUND: In the present study, an activity of Bixa orellana extract against Leishmania amazonensis was demonstrated. RESULT: Experimentally infected BALB/c mice were treated with B. orellana extract which showed a significant activity against promastigote and amastigote forms of L. amazonensis. CONCLUSION: This study supports the importance of natural sources as antileishmanial drugs.

Asunto(s)

Antiprotozoarios/farmacología , Antiprotozoarios/uso terapéutico , Bixaceae/química , Leishmania/efectos de los fármacos , Leishmaniasis/tratamiento farmacológico , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Anfotericina B/farmacología , Animales , Peso Corporal/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Femenino , Concentración 50 Inhibidora , Estadios del Ciclo de Vida/efectos de los fármacos , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/parasitología , Ratones , Ratones Endogámicos BALB C , Distribución Aleatoria

RESUMEN

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Asunto(s)

Variación Genética/genética , Macrófagos Peritoneales/parasitología , Óxido Nítrico/biosíntesis , Fagocitosis/fisiología , Trypanosoma cruzi/genética , Animales , Reservorios de Enfermedades/parasitología , Femenino , Interacciones Huésped-Parásitos , Humanos , Insectos Vectores/parasitología , Macrófagos Peritoneales/citología , Ratones , Ratones Endogámicos BALB C , Triatominae/parasitología , Trypanosoma cruzi/clasificación , Trypanosoma cruzi/fisiología

RESUMEN

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

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Doze cepas de Trypanosoma cruzi isoladas de reservatórios silvestres, triatomíneos e de pacientes chagásicos crônicos do Estado do Paraná, Brasil, classificadas como Tc I e II foram usadas para avaliar a correlação entre genética e diversidade biológica. Índice fagocítico (IF) e produção de óxido nítrico (ON) in vitro foram os parâmetros biológicos utilizados. O IF de cepas T. cruzi I e II não diferiram significativamente assim como o IF de cepas isoladas de humanos, triatomíneos ou de reservatórios silvestres. Há diferença estatística na inibição da produção de ON entre T. cruzi I e II e entre parasitos isolados de humanos e de cepas isoladas de triatomíneos e reservatórios silvestres, mas não foi observada correlação entre genética e biologia quando as cepas foram analisadas independentemente da linhagem ou hospedeiros das quais elas foram isoladas. Observou-se correlação significativa para amplificação aleatória do DNA polimórfico e parâmetros biológicos de Tc I ou II e para os seres humanos ou reservatório silvestre quando linhagens ou hospedeiros são consideradas separadamente.

Asunto(s)

Animales , Femenino , Humanos , Ratones , Variación Genética/genética , Macrófagos Peritoneales/parasitología , Óxido Nítrico/biosíntesis , Fagocitosis/fisiología , Trypanosoma cruzi/genética , Reservorios de Enfermedades/parasitología , Interacciones Huésped-Parásitos , Insectos Vectores/parasitología , Ratones Endogámicos BALB C , Macrófagos Peritoneales/citología , Triatominae/parasitología , Trypanosoma cruzi/clasificación , Trypanosoma cruzi/fisiología