ABSTRACT
AIM: To evaluate the cytotoxicity and the mechanism of cell aggression of peracetic acid (PA) in comparison with sodium hypochlorite (NaOCl). METHODOLOGY: L929 fibroblasts were exposed to 1% PA and 2.5% NaOCl, at several dilutions for 10 min. The following parameters were evaluated: cell metabolism by methylthiazol tetrazolium assay, external morphology by scanning electron microscopy, ultrastructure by transmission electron microscopy, the cytoskeleton by means of actin and α-tubulin labelling, and the type of cell death by flow cytometry (apoptosis/necrosis). The data were analysed by two-way anova and the Bonferroni post-test (α = 0.05). RESULTS: The PA group had lower cell viability and a higher percentage of necrotic cells than the NaOCl group (P < 0.05). Both solutions diminished cell metabolism, led to destructuring of the cytoskeleton, created changes in the external morphology, resulted in the accumulation of proteins in the rough endoplasmic reticulum and induced cell death predominantly by necrosis. However, these changes were observed in lower doses of PA when compared with NaOCl. CONCLUSIONS: Although they had the same mechanism of cytotoxicity, 1% PA had greater cytotoxic potential than 2.5% NaOCl.
Subject(s)
Apoptosis/drug effects , Disinfectants/toxicity , Fibroblasts/drug effects , Peracetic Acid/toxicity , Animals , Cell Culture Techniques , Cell Line , Cell Survival/drug effects , Cytoskeleton/drug effects , Fibroblasts/metabolism , Fibroblasts/ultrastructure , Flow Cytometry , Mice , Microscopy, Electron , Necrosis , Sodium Hypochlorite/toxicityABSTRACT
Abstract Garlic (Allium sativum L.) is grown all over the world as seasoning and medicinal vegetable since 3,000 BC. Allicin is the main component of garlic, being attributed to it the most of its biological activities, such as bactericidal, antifungal and antiviral actions. However, other compounds of garlic present antioxidant, hypocholesterolemic, vasodilator activities, protective action against different types of cancer, and immunomodulatory. Fungal infections are important causes of morbidity and mortality in people mainly in immunosuppressed ones. Sporothrix schenckii, the causing agent of Sporotrichosis (most common subcutaneous mycosis in Latin America), is dimorphic fungus, of saprophytic life in soil or plants, infecting people and animals mainly through skin injuries and bruises. The main of this work was to evaluate the influence of garlic consuming on immune modulation of healthy and infected Swiss mice in induced way by S. schenckii, since these animals functioning of peritoneal macrophages as well as the nitric oxide and cytokines' production (IL-1β, IL-10 and IL-12) and to evaluate the antifungal potential of garlic with S. schenckii through minimum inhibitory concentration test and colony-forming units. The results showed that garlic offers antifungal potential with S. schenckii. The oral taking of garlic extracts influences the releasing of cytokines by macrophages, regular consuming shows anti-inflammatory effect, and its acute use may take to an inflammatory response. Mice that consumed garlic responded more effectively to fight against the infection.
Resumo O alho (Allium sativum L.) é cultivado em todo o mundo como hortaliça condimentar e medicinal desde 3.000 a. C. A alicina é o principal componente do alho, sendo atribuída a ela a maior parte das suas atividades biológicas, dentre elas as ações bactericida, antifúngica e antiviral. Porém, outros compostos do alho apresentam atividade antioxidante, hipocolesterolemiante, vasodilatadora, ação protetora contra diversos tipos de câncer e imunomoduladora. As infecções por fungos são causas importantes de morbidade e mortalidade no homem principalmente em indivíduos imunossuprimidos. O Sporothrix schenckii, agente causal da esporotricose (micose subcutânea mais comum na América Latina), é fungo dimórfico, de vida saprofítica no solo ou em vegetais, infectando homens e os animais principalmente através de lesões e arranhões na pele. O objetivo desse trabalho foi avaliar a influência do consumo de alho na imunomodulação de camundongos Swiss saudáveis e infectados de forma induzida por S. schenckii, a partir do estado funcional dos macrófagos peritoneais desses animais quanto à produção de óxido nítrico e das citocinas (IL-1β, IL-10 e IL-12) e avaliar o potencial antifúngico do alho frente ao S. schenckii por meio de teste de concentração inibitória mínima e unidades formadoras de colônia. Os resultados demonstraram que o alho apresenta potencial antifúngico frente S. schenckii. A administração oral de extratos de alho influencia a liberação de citocinas por macrófagos, o consumo regular apresenta efeito anti-inflamatório, e seu uso agudo pode gerar uma resposta inflamatória. Camundongos que consumiram alho responderam de forma mais efetiva no combate da infecção.
Subject(s)
Animals , Male , Mice , Sporotrichosis/drug therapy , Sporothrix/drug effects , Plant Extracts/pharmacology , Macrophages, Peritoneal/immunology , Garlic/chemistry , Antifungal Agents/pharmacology , Cytokines/metabolism , Macrophages, Peritoneal/drug effects , ImmunomodulationABSTRACT
Garlic (Allium sativum L.) is grown all over the world as seasoning and medicinal vegetable since 3,000 BC. Allicin is the main component of garlic, being attributed to it the most of its biological activities, such as bactericidal, antifungal and antiviral actions. However, other compounds of garlic present antioxidant, hypocholesterolemic, vasodilator activities, protective action against different types of cancer, and immunomodulatory. Fungal infections are important causes of morbidity and mortality in people mainly in immunosuppressed ones. Sporothrix schenckii, the causing agent of Sporotrichosis (most common subcutaneous mycosis in Latin America), is dimorphic fungus, of saprophytic life in soil or plants, infecting people and animals mainly through skin injuries and bruises. The main of this work was to evaluate the influence of garlic consuming on immune modulation of healthy and infected Swiss mice in induced way by S. schenckii, since these animals functioning of peritoneal macrophages as well as the nitric oxide and cytokines' production (IL-1ß, IL-10 and IL-12) and to evaluate the antifungal potential of garlic with S. schenckii through minimum inhibitory concentration test and colony-forming units. The results showed that garlic offers antifungal potential with S. schenckii. The oral taking of garlic extracts influences the releasing of cytokines by macrophages, regular consuming shows anti-inflammatory effect, and its acute use may take to an inflammatory response. Mice that consumed garlic responded more effectively to fight against the infection.
Subject(s)
Antifungal Agents/pharmacology , Garlic/chemistry , Macrophages, Peritoneal/immunology , Plant Extracts/pharmacology , Sporothrix/drug effects , Sporotrichosis/drug therapy , Animals , Cytokines/metabolism , Immunomodulation , Macrophages, Peritoneal/drug effects , Male , MiceABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Vismia guianensis (Aubl.) Pers. is traditionally used in North and Northeast of Brazil for the treatment of dermatomycoses. Since the strategy associating immunomodulators with antifungal drugs seems to be promissory to improve the treatment efficacy in fungal infections, we aimed to investigate the antifungal activity of V. guianensis ethanolic extract of leaves (VGL) and bark (VGB) against Sporothrix schenckii ATCC 16345 and their antinflammatory activities. MATERIAL AND METHODS: The extracts were analyzed by HPLC-DAD-IT MS/MS for in situ identification of major compounds. Antifungal activity was evaluated in vitro (microdilution test) and in vivo using a murine model of S. schenckii infection. The production of TNF-α, IFN-γ, IL-4, IL-10 and IL-12 by measured by ELISA, as well as measured the production and inhibition of the NO after treatment with the plant extracts or itraconazole (ITR). RESULTS: Two O-glucosyl-flavonoids and 16 prenylated benzophenone derivatives already described for Vismia were detected. Both VGL and VGB showed significant antifungal activity either in in vitro assay of microdilution (MIC=3.9µg/mL) and in vivo model of infection with reduction of S. schenckii load in spleen. It was also observed a predominance of reduction in the production of NO and the proinflammatory cytokines evaluated except TNFα, but with stimulation of IL-10, as evidence of a potential anti-inflammatory effect associated. CONCLUSION: The results showed that both VGL and VGB have a significant antifungal against S. schenckii and an anti-inflammatory activity. These results can support the use of these extracts for alternative treatment of sporotrichosis.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Antifungal Agents/pharmacology , Clusiaceae/chemistry , Plant Extracts/pharmacology , Sporothrix/drug effects , Sporotrichosis/prevention & control , Animals , Anti-Inflammatory Agents/isolation & purification , Antifungal Agents/isolation & purification , Cells, Cultured , Chromatography, High Pressure Liquid , Disease Models, Animal , Ethanol/chemistry , Interferon-gamma/metabolism , Interleukin-10/metabolism , Interleukin-12/metabolism , Interleukin-4/metabolism , Itraconazole/pharmacology , Male , Mice, Inbred BALB C , Nitric Oxide/metabolism , Phytotherapy , Plant Bark/chemistry , Plant Extracts/isolation & purification , Plant Leaves/chemistry , Plants, Medicinal , Solvents/chemistry , Sporothrix/immunology , Sporotrichosis/immunology , Sporotrichosis/metabolism , Sporotrichosis/microbiology , Tandem Mass Spectrometry , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Introduction: Sporotrichosis is an emergent subcutaneous mycoses caused by species of the Sporothrix schenckii complex. Amphotericin B (AmB) remains the main antifungal drug for the treatment of systemic infections, but its use is limited by toxicity reasons. AFCo3 is a novel cochleate containing detoxified LPS, which exhibits drug delivery and immunomodulating properties. Here, AFCo3 was used as the vehicle for AmB to evaluate the immunomodulatory and antifungal efficacy against S. schenckii in vitro and in vivo. Methods and results: The minimum inhibitory concentrations of AFCo3-AmB and AmB were 0.25 and 1 μg/mL respectively. The minimum fungicidal concentration was 0.5 μg/mL for AFCo3-AmB and 2 μg/mL for AmB. AFCo3-AmB was less cytotoxic than AmB for peritoneal macrophages, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and reduced the AmB-induced hemolysis in murine erythrocytes. AFCo3-AmB improved the intracellular killing of phagocytized yeast and it enhanced the in vitro production of IL-1β, TNF-α and NO in peritoneal macrophages. Moreover, AFCo3-AmB was more effective than AmB in reducing spleen and liver fungal burden after repeated (five days) intraperitoneal administration of 5 mg/kg of AmB, in a Balb/c model of systemic infection, associated to a significant induction of Th1/Th17 response. Finally, blood chemistry revealed that AFCo3-AmB did not cause changes suggestive of nephrotoxicity, such as increases in total proteins, albumin, creatinine and blood urea nitrogen that were caused by free AmB. Conclusions: AFCo3-AmB exhibited asignificant immunomodulator action, reduced toxicity and improved antifungal action against S. schenckii, suggesting a potential use as AmB delivery for systemic sporotrichosis treatment(AU)
Subject(s)
Humans , Sporotrichosis , Antifungal Agents/toxicity , Amphotericin B/therapeutic use , Mycosis Fungoides/therapy , Immunomodulation , SporothrixABSTRACT
INTRODUCTION: Sporotrichosis is an emergent subcutaneous mycoses caused by species of the Sporothrix schenckii complex. Amphotericin B (AmB) remains the main antifungal drug for the treatment of systemic infections, but its use is limited by toxicity reasons. AFCo3 is a novel cochleate containing detoxified LPS, which exhibits drug delivery and immunomodulating properties. Here, AFCo3 was used as the vehicle for AmB to evaluate the immunomodulatory and antifungal efficacy against S. schenckii in vitro and in vivo. METHODS AND RESULTS: The minimum inhibitory concentrations of AFCo3-AmB and AmB were 0.25 and 1µg/mL respectively. The minimum fungicidal concentration was 0.5µg/mL for AFCo3-AmB and 2µg/mL for AmB. AFCo3-AmB was less cytotoxic than AmB for peritoneal macrophages, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and reduced the AmB-induced hemolysis in murine erythrocytes. AFCo3-AmB improved the intracellular killing of phagocytized yeast and it enhanced the in vitro production of IL-1ß, TNF-α and NO in peritoneal macrophages. Moreover, AFCo3-AmB was more effective than AmB in reducing spleen and liver fungal burden after repeated (five days) intraperitoneal administration of 5mg/kg of AmB, in a Balb/c model of systemic infection, associated to a significant induction of Th1/Th17 response. Finally, blood chemistry revealed that AFCo3-AmB did not cause changes suggestive of nephrotoxicity, such as increases in total proteins, albumin, creatinine and blood urea nitrogen that were caused by free AmB. CONCLUSIONS: AFCo3-AmB exhibited a significant immunomodulator action, reduced toxicity and improved antifungal action against S. schenckii, suggesting a potential use as AmB delivery for systemic sporotrichosis treatment.
Subject(s)
Amphotericin B/administration & dosage , Antifungal Agents/administration & dosage , Drug Carriers/administration & dosage , Immunologic Factors/administration & dosage , Lipopolysaccharides/administration & dosage , Macrophages, Peritoneal/drug effects , Sporothrix/drug effects , Amphotericin B/pharmacology , Amphotericin B/therapeutic use , Animals , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Cell Survival/drug effects , Cytokines/metabolism , Drug Carriers/pharmacology , Drug Carriers/therapeutic use , Erythrocytes/drug effects , Hemolysis/drug effects , Immunologic Factors/pharmacology , Immunologic Factors/therapeutic use , Lipopolysaccharides/pharmacology , Lipopolysaccharides/therapeutic use , Liver/drug effects , Liver/microbiology , Macrophages, Peritoneal/metabolism , Macrophages, Peritoneal/microbiology , Male , Mice, Inbred BALB C , Microbial Sensitivity Tests , Nitric Oxide/metabolism , Spleen/cytology , Spleen/drug effects , Spleen/microbiology , Sporothrix/growth & development , Sporotrichosis/drug therapy , Sporotrichosis/microbiologyABSTRACT
Sporothrix schenckii is a complex of various species of fungus found in soils, plants, decaying vegetables and other outdoor environments. It is the aetiological agent of sporotrichosis in humans and several animals. Humans and animals can acquire the disease through traumatic inoculation of the fungus into subcutaneous tissue. Despite the importance of sporotrichosis, it being currently regarded as an emergent disease in several countries, the factors driving its increasing medical importance are still largely unknown. There have only been a few studies addressing the influence of the environment on the virulence of these pathogens. However, recent studies have demonstrated that adverse conditions in its natural habitats can trigger the expression of different virulence factors that confer survival advantages both in animal hosts and in the environment. In this review, we provide updates on the important advances in the understanding of the biology of Spor. schenckii and the modification of its virulence linked to demonstrated or putative environmental factors.
Subject(s)
Sporothrix/pathogenicity , Sporotrichosis/microbiology , Animals , Environmental Microbiology , Fungal Proteins/genetics , Fungal Proteins/metabolism , Humans , Sporothrix/genetics , Sporothrix/physiology , VirulenceABSTRACT
Sporotrichosis is a disease caused by the dimorphic fungus Sporothrix schenckii. The main clinical manifestations occur in the skin, however the number of systemic and visceral cases has increased, especially in immunocompromised patients. Dendritic cells (DCs) are highly capable to recognize the fungus associated data and translate it into differential T cells responses both in vivo and in vitro. Although, the mechanisms involved in the interaction between DCs and S. schenckii are not fully elucidated. The present study investigated the phenotypic and functional changes in bone marrow dendritic cells (BMDCs) stimulated in vitro with the yeast form of S. schenckii or exoantigen (ExoAg) and its ability to trigger a cellular immune response in vitro. Our results demonstrated that the live yeast of S. schenckii and its exoantigen, at a higher dose, were able to activate BMDCs and made them capable of triggering T cell responses in vitro. Whereas the yeast group promoted more pronounced IFN-γ production rather than IL-17, the Exo100 group generated similar production of both cytokines. The exoantigen stimulus suggests a capability to deviate the immune response from an effector Th1 to an inflammatory Th17 response. Interestingly, only the Exo100 group promoted the production of IL-6 and a significant increase of TGF-ß, in addition to IL-23 production. Interestingly, only Exo100 group was capable to promote the production of IL-6 and a significant increase on TGF-ß, in addition with IL-23 detection. Our results demonstrated the plasticity of DCs in translating the data associated with the fungus S. schenckii and ExoAg into differential T cell responses in vitro. The possibility of using ex vivo-generated DCs as vaccinal and therapeutic tools for sporotrichosis is a challenge for the future.
Subject(s)
Antigens, Fungal/immunology , Dendritic Cells/immunology , Sporothrix/immunology , Sporotrichosis/immunology , Th1 Cells/immunology , Th17 Cells/immunology , Animals , Bone Marrow Cells/immunology , Bone Marrow Cells/metabolism , CD11c Antigen/immunology , CD11c Antigen/metabolism , Cell Differentiation/immunology , Cells, Cultured , Coculture Techniques , Cytokines/immunology , Cytokines/metabolism , Dendritic Cells/metabolism , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Host-Pathogen Interactions/immunology , Humans , Interleukin-23/immunology , Interleukin-23/metabolism , Interleukin-6/immunology , Interleukin-6/metabolism , Lymphocyte Activation/immunology , Mice , Mice, Inbred C57BL , Sporothrix/physiology , Sporotrichosis/metabolism , Sporotrichosis/microbiology , Th1 Cells/metabolism , Th17 Cells/metabolism , Transforming Growth Factor beta/immunology , Transforming Growth Factor beta/metabolismABSTRACT
Dois biomarcadores do estresse oxidativo foram avaliados em indivíduos saudáveis que receberam doses diárias de suco de laranja, notadamente uma fonte de vitamina C e de flavanonas, que têm sido associadas aos efeitos antioxidante, anti-inflamatório e hipolipidêmico. A capacidade antioxidante do soro sanguíneo foi avaliada através da redução do radical 2,2-difenil-1-picril-hidrazila (DPPH) e a peroxidação lipídica foi avaliada pela presença de substâncias reativas ao ácido tiobarbitúrico (TBARS). Os resultados mostraram que após o período de suplementação com o suco de laranja houve um aumento 150% na capacidade antioxidante no soro das mulheres e 200% no soro dos homens (p<0,001), mas não houve alteração na peroxidação lipídica no sangue dos voluntários. Concluindo, o aumento das reservas de flavanonas e de vitamina C, decorrente da ingestão regular de suco de laranja, melhorou expressivamente a capacidade antioxidante no sangue, sem, entretanto, apresentar efeito sobre a peroxidação lipídica.
Two biomarkers of oxidative stress were evaluated in healthy volunteers treated with daily doses of orange juice that is well known source of vitamin C and citric flavanones, which have been associated with antioxidant, anti-inflammatory and hypolipidemic effects. The antioxidant capacity in the blood serum was evaluated by studying the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical, and also lipid peroxidation was evaluated by the thiobarbituric acid reacting substances (TBARS) assay. The results have shown that the regular consumption of orange juice increased 150% and 200% the serum antioxidant capacity for women and men respectively, but it was no significant change in the serum lipid peroxidation. In conclusion, the increase of flavonones and vitamin C in the body due to the regular intake of orange juice expressively improved the antioxidant capacity, but without significant effect on the lipid peroxidation.
Subject(s)
Humans , Ascorbic Acid/blood , Carbonated Beverages , Citrus sinensis , Flavones/blood , Lipid Peroxidation , Oxidative StressABSTRACT
Complexes of the type [PdX2(tdmPz)] {X=Cl-(1), Br-(2); I-(3); SCN-(4); tdmPz=1-thiocarbamoyl-3,5-dimethylpyrazole} have been synthesized and characterized. Compound 1 was formed from the reaction between [PdCl2(CH3CN)2] and 1-thiocarbamoyl-3,5-dimethylpyrazole. Complexes 2, 3 and 4 were obtained by metathesis of the chloro groups from 1 by bromide, iodide and thiocyanate ions, respectively. All the compounds and cisplatin have been tested in vitro by MTT assay for their cytotoxicity against three murine cancer cell lines: mammary adenocarcinoma (LM3 and LMM3) and lung adenocarcinoma (LP07) as well towards normal murine peritoneal exudate cells (PEC). Promising cytotoxic effect against LM3 has been found for 3 showing IC50 equal to 24.5 microM which is comparable to the value obtained for cisplatin (30.3 microM).
Subject(s)
Antineoplastic Agents/pharmacology , Organometallic Compounds/pharmacology , Palladium/chemistry , Pyrazoles/chemistry , Animals , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Cell Line, Tumor , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Female , Macrophages, Peritoneal/drug effects , Magnetic Resonance Spectroscopy , Mice , Mice, Inbred BALB C , Molecular Structure , Organometallic Compounds/chemical synthesis , Organometallic Compounds/chemistry , Spectrophotometry, Infrared , Spectrophotometry, Ultraviolet , Stereoisomerism , Structure-Activity RelationshipABSTRACT
Plantas têm contribuído no tratamento da maioria das doenças. Considerando a importância terapêutica das plantas medicinais, foi avaliada a atividade imunológica dos extratos metanólico e etanólico de Davilla elliptica. Macrófagos estão envolvidos em todos estágios da resposta imune, podendo liberar componentes como: peróxido de hidrogênio (H2O2), óxido nítrico (NO) e fator de necrose tumoral-alfa (TNF-alfa). A estimulação dos macrófagos foi avaliada pela determinação de H2O2, NO e TNF-alfa em culturas de macrófagos peritoneais de camundongos na presença dos extratos da D. elliptica. IC50 foi determinado através de ensaio utilizando MTT. Os estudos fitoquímicos realizados mostraram a presença de flavonóides derivados da quercetina e miricetina entre outros compostos. A produção de H2O2 não foi muito expressiva em ambos extratos, contudo a de NO foi significativa. Os dois extratos induziram a produção de TNF-alfa, sendo que a liberação dessa citocina pelo extrato metanólico foi quase cinco vezes maior do que pelo extrato etanólico. Uma relação entre as sínteses de NO e TNF-alfa foi observada. O aumento na produção de NO está relacionado com a indução de citocinas pró-inflamatórias como TNF-alfa. Analisando os resultados, sugere-se que os extratos metanólico e etanólico de D. elliptica podem modular a ativação de macrófagos.
Plants have contributed in a significant way to the treatment of most of the diseases. Considering the therapeutic importance of the medicinal plants, the immunological activity of the methanolic and ethanolic extract of Davilla elliptica was evaluated. In every stage of the immune response macrophages are involved and they can release many compounds such as hydrogen peroxide (H2O2), nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha). Macrophages stimulation was evaluated by the determination of H2O2, NO and TNF-alpha in peritoneal macrophages cultures of mice in the presence of the D. elliptica extracts. IC50 was determined by MTT assay. The phytochemical study showed flavonoids derived from quercetin and myricetin and other compounds. The production of H2O2 was not very expressive in both extracts, but they presented a significant effect on NO production. The two extracts induced TNF-alpha production, although the methanolic liberated almost five times more TNF-alpha than the ethanolic one. A relationship among the synthesis of NO and TNF-alpha was observed. The increase of NO production is related with the induction of proinflammatory cytokines like TNF-alpha. Analyzing the results, it is suggested that methanolic and ethanolic extract of D. elliptica can modulate macrophage activation.
ABSTRACT
The activity of ten compounds isolated from Brazilian lichen over the release of hydrogen peroxide and nitric oxide was evaluated in the culture of peritoneal macrophage cells from mice. Salazinic, secalonic A and fumarprotocetraric acids were the compounds that induced the greatest release of H2O2, whereas 12R-usnic and diffractaic acids induced the release of NO. These results indicate that lichen products have potential immunological modulating activities.
Subject(s)
Biological Factors/pharmacology , Lichens , Macrophages, Peritoneal/drug effects , Phytotherapy , Plant Extracts/pharmacology , Animals , Biological Factors/administration & dosage , Biological Factors/therapeutic use , Brazil , Hydrogen Peroxide/metabolism , Macrophages, Peritoneal/metabolism , Male , Medicine, Traditional , Mice , Nitric Oxide/metabolism , Plant Extracts/administration & dosage , Plant Extracts/therapeutic useABSTRACT
Chlorhexidine, even at low concentrations, is toxic for a variety of eukaryotic cells; however, its effects on host immune cells are not well known. We evaluated in vitro chlorhexidine-induced cytotoxicity and its effects on reactive oxygen/nitrogen intermediate induction by murine peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 ml of 10 mM phosphate-buffered saline, washed twice and resuspended (10(6) cells/ml) in appropriate medium for each test. Cell preparations contained more than 95% macrophages. The cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the horseradish peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. The midpoint cytotoxicity values for 1- and 24-h exposures were 61.12 +/- 2.46 and 21.22 +/- 2.44 microg/ml, respectively. Chlorhexidine did not induce synthesis or liberation of reactive oxygen/nitrogen intermediates. When macrophages were treated with various sub-toxic doses for 1 h (1, 5, 10, and 20 microg/ml) and 24 h (0.5, 1, and 5 microg/ml) and stimulated with 200 nM phorbol myristate acetate (PMA) solution, the H2O2 production was not altered; however, the NO production induced by 10 microg/ml lipopolysaccharide (LPS) solution varied from 14.47 +/- 1.46 to 22.35 +/- 1.94 micromol/l and 13.50 +/- 1.42 to 20.44 +/- 1.40 micromol/l (N = 5). The results showed that chlorhexidine has no immunostimulating activity and sub-toxic concentrations did not affect the response of macrophages to the soluble stimulus PMA but can interfere with the receptor-dependent stimulus LPS.
Subject(s)
Anti-Infective Agents/toxicity , Chlorhexidine/analogs & derivatives , Chlorhexidine/toxicity , Hydrogen Peroxide/metabolism , Macrophages, Peritoneal/drug effects , Nitric Oxide/biosynthesis , Animals , Cell Survival/drug effects , Lipopolysaccharides/pharmacology , Mice , Reactive Nitrogen Species/metabolism , Reactive Oxygen Species/metabolism , Tetradecanoylphorbol Acetate/pharmacologyABSTRACT
Chlorhexidine, even at low concentrations, is toxic for a variety of eukaryotic cells; however, its effects on host immune cells are not well known. We evaluated in vitro chlorhexidine-induced cytotoxicity and its effects on reactive oxygen/nitrogen intermediate induction by murine peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 ml of 10 mM phosphate-buffered saline, washed twice and resuspended (10(6) cells/ml) in appropriate medium for each test. Cell preparations contained more than 95 percent macrophages. The cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the horseradish peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. The midpoint cytotoxicity values for 1- and 24-h exposures were 61.12 ± 2.46 and 21.22 ± 2.44 æg/ml, respectively. Chlorhexidine did not induce synthesis or liberation of reactive oxygen/nitrogen intermediates. When macrophages were treated with various sub-toxic doses for 1 h (1, 5, 10, and 20 æg/ml) and 24 h (0.5, 1, and 5 æg/ml) and stimulated with 200 nM phorbol myristate acetate (PMA) solution, the H2O2 production was not altered; however, the NO production induced by 10 æg/ml lipopolysaccharide (LPS) solution varied from 14.47 ± 1.46 to 22.35 ± 1.94 æmol/l and 13.50 ± 1.42 to 20.44 ± 1.40 æmol/l (N = 5). The results showed that chlorhexidine has no immunostimulating activity and sub-toxic concentrations did not affect the response of macrophages to the soluble stimulus PMA but can interfere with the receptor-dependent stimulus LPS.
Subject(s)
Animals , Mice , Anti-Infective Agents , Chlorhexidine , Hydrogen Peroxide , Macrophages, Peritoneal , Nitric Oxide , Lipopolysaccharides , Reactive Oxygen Species , Tetradecanoylphorbol AcetateABSTRACT
A resposta imunológica engloba contexto amplo envolvendo várias células, sendo o macrófago importante na resposta inflamatória que antecede a resposta imune celular. Existem vários estímulos à membrana macrofágica que podem desencadear a liberação de H2O2, contribuindo para uma ação antimicrobiana e citotóxica. Atualmente há uma forte tendência em se estudar produtos naturais quanto à capacidade de atuação no sistema imunológico. Este estudo avaliou a citotoxicidade do extrato bruto (EB) e das frações hexânica (HX) e acética (AC) extraídos do caule de Styrax camporum Pohl (Styracaceae) e determinou a concentração de H2O2 nos sobrenadantes de culturas de macrófagos peritoneais de camundongos expostos à EB, HX e AC nas concentrações de 4,2 e 1 mg/ml. A fração HX a 2 mg/ml foi capaz de promover significativa liberação de H2O2, quando comparada com Zimosan (controle positivo). Na concentração de 4 mg/ml, HX e EB apresentaram alta toxicidade. A fração AC não promoveu nenhuma liberação de H2O2, enquanto EB produziu baixa liberação. Concluiu-se que a fração HX é um potente estimulador de macrófagos, apresentando toxicidade elevada em altas concentrações
Subject(s)
Animals , Male , Mice , Macrophages, Peritoneal , Plants, Medicinal/toxicity , Plant Extracts/toxicity , Hydrogen PeroxideABSTRACT
The role of cell-wall compounds in the immune response to sporotrichosis is unknown. The effect of cell-wall compounds and exoantigen obtained from Sporothrix schenckii in macrophage/fungus interaction was analysed with respect to nitric oxide (NO) and tumour necrosis factor-alpha (TNF-alpha). The lipid compound of the cell wall plays an important role in the pathogenesis of this mycosis and was found to inhibit the phagocytic process and to induce high liberation of NO and TNF-alpha in macrophage cultures in the present study. This is a very interesting result because it is the first report about one compound of the fungus S. schenckii that presents this activity.
Subject(s)
Lipids/immunology , Macrophages, Peritoneal/immunology , Nitric Oxide/immunology , Sporothrix/immunology , Tumor Necrosis Factor-alpha/immunology , Animals , Antigens, Fungal/immunology , Cell Wall/immunology , Hydrogen Peroxide/metabolism , Lipids/pharmacology , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/metabolism , Mice , Nitric Oxide/metabolism , Nitrites/metabolism , Phagocytosis/drug effects , Phagocytosis/immunology , Sporotrichosis/immunology , Tumor Necrosis Factor-alpha/metabolismABSTRACT
This study was undertaken to verify the effect of a daily intake of a new fermented soy milk produced with Enterococcus faecium and Lactobacillus jugurti on the serum lipid levels in normocholesterolemic middle-aged men. The study was randomized, double-blind and placebo-controlled and was performed for a period of 6 weeks. Forty-four normocholesterolemic healthy, male volunteers, aged 40-55 years old were randomly separated in two groups: The F-group received 200 ml of the fermented product daily and the P-group received 200 ml of placebo (chemically fermented). The blood samples were drawn initially and after 3 and 6 weeks and serum values for total cholesterol, HDL-cholesterol and triglyceride were determined. The LDL-cholesterol value was estimated. No significant changes in the fermented group (F) were observed for total cholesterol, LDL-cholesterol or triglyceride levels, while the HDL-cholesterol level was significantly higher (p < or = 0.05) after 6 weeks. The total cholesterol and LDL-cholesterol levels were significantly higher (p < or = 0.05) in the placebo group (P), but no changes were found for the HDL-cholesterol and triglyceride levels during the experimental period. In conclusion, the intake of 200 ml/day of the fermented soy milk, produced with E. faecium and L. jugurti, for 6 weeks, did not affect the serum total cholesterol and LDL-cholesterol, and led an increase of 10 per cent in the HDL-cholesterol level.
Subject(s)
Humans , Male , Adult , Middle Aged , Cholesterol/blood , Glycine max , Yogurt , Arteriosclerosis/etiology , Arteriosclerosis/prevention & control , Cholesterol, HDL , Cholesterol, LDL , Cholesterol/metabolism , Double-Blind Method , Eating , Enterococcus faecium , Fermentation , Lactobacillus , Lipids/blood , Lipids/metabolism , Risk Factors , Triglycerides/bloodABSTRACT
Macrófagos são as primeiras células a participarem da resposta imunológica, e quando são ativados liberam mais de cem compostos ao meio extracelular, entre os compostos reativos intermediários de nitrogênio (NO). Neste trabalho determinou-se a liberação de óxido nítrico em culturas de macrófagos peritoneais de camundongos em presença de óleo essencial bruto e extrato etanólico 70 por cento bruto obtidos a partir de folhas de Achillea millefolim L. (Asteraceae). Diferentes diluições do óleo essencial foram testadas (1:50, 1:100 e 1:200). Apenas a diluição 1:100 produziu uma maior quantidade de óxido nítrico (NO). Em relação ao extrato etanólico 70 por cento, observou-se nas amostras mais concentradas (6 mg/mL, 8 mg/mL e 10 mg/mL) maior produção de NO. Analisando-se os resultados obtidos no presente trabalho, pode-se sugerir que tanto o óleo essencial quanto o extrato etanólico 70 por cento bruto de A. millefolium L são agentes moduladores da ativação de macrófagos, nas concentrações de 20, 10 e 5 mg/mL, quando comparado com LPS (lipopolissacarídeo-potente estimulador da produção de NO).
ABSTRACT
As plantas têm sido valiosa fonte de produtos naturais para a manutenção da saúde humana. O objetivo deste trabalho foi estudar a atividade imunológica da mil-folhas (Achillea millefolium L.), planta muito utilizada na medicina popular por possuir uma série de ações, como, por exemplo, antiinflamatória, adstringente, anti-séptica e antiespasmódica. Avaliou-se a ativação de macrófagos, determinando-se a liberação de óxido nítrico (NO), peróxido de hidrogênio (N2O2) e do fator de necrose tumoral-alfa (TNF-alfa) em sobrenadantes de culturas de macrófagos peritoneais de camundongos, na presença do extrato etanólico 70 por cento bruto da mil-folhas. A análise cromatográfica do extrato também foi realizada, revelando presença de rutina. Observou-se liberação moderada de H2O2 em todas as concentrações de extrato testadas, enquanto que somente nas amostras mais concentradas (6,8 e 10mg/mL) ocorreu maior produção de NO. Houve moderada produção de TNT-a em todas as amostras, sendo que a maior ocorreu na concentração de 4mg/mL. Pelos resultados encontrados, sugere-se que o extrato da mil-folhas é um agente modulador da ativação de macrófagos.