RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Wound healing is a complex process that can leave pathological scars, especially in case of infections from opportunistic microorganisms. In this context, herbal medicines open up great possibilities for investigation. One of the species of interest native to Brazil is Garcinia brasiliensis ("bacupari"). Traditionally known for treating wounds and ulcers, G. brasiliensis presents anti-inflammatory, antioxidant and antimicrobials properties. But, its wound healing profile in experimental models, in order to validate its efficacy, is still litle studied. AIM OF THE STUDY: Thus, the objective of this work was to evaluate, in an infected cutanous wound model, the potential of formulations incorporated with G. brasiliensis leaves extracts. MATERIALS AND METHODS: Crude extract (CE), Ethyl Acetate Fraction (EAF) and Hexanic Fraction (HF) were submitted to phytochemical assays, high performance thin layer chromatography (HTPLC) and cytotoxicity studies. CE and EAF were also tested for microbicidal properties and incorporated in cream and gel formulations at 10% concentration. After stability testing, the gel formulations with CE or EAF at 10% were selected and applied to skin wounds infected or not with Staphylococcus aureus in Wistar rats. The healing potenttial of the extracts was verified by the expression of the protein Annexin A1 (AnxA1), related to the processes of inflammation and antifibrotic function, the cells immunostaining for Gasdermin-D (GSDM-D), a marker of pyroptotic cell death, and the dosage of interleukin-10 (IL-10) and monocyte chemotactic protein (MCP)-1 inflammatory mediators. RESULTS: Phytochemical studies indicated the presence of compounds of pharmacological interest, including Catechin, Quercetin and Berberine in addition to low cytotoxicity of CE and EAF at 10%. After the 6-day topical treatments, CE and EAF gel formulations demonstrated to control the pruritus formation process. The treatments decreased AnxA1 expression and the amount of cells immunostained for GSDM-D, and increased the expression of MCP-1 in infected wounds. CONCLUSIONS: Together, the results show important anti-inflammatory profile and skin healing potential of CE and EAF from G. brasiliensis leaves, even in infected lesions, with therapeutic perspectives.
Assuntos
Garcinia , Animais , Anti-Inflamatórios/análise , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Compostos Fitoquímicos/análise , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Folhas de Planta/química , Ratos , Ratos Wistar , CicatrizaçãoRESUMO
Introdução: O benzopireno é um dos principais hidrocarbonetos aromáticos policíclicos presentes no ambiente e com alta capacidade carcinogênica, sendo, portanto, usado em modelos in vivo de carcinogênese pulmonar. Investigações têm mostrado o envolvimento dos mastócitos na modulação do ambiente tumoral e que os fármacos anti-inflamatórios podem reduzir a incidência de câncer de pulmão. Entre as possibilidades terapêuticas estão os fitoterápicos. O extrato de Garcinia brasiliensis, conhecido popularmente como bacupari, mostra propriedades anti-inflamatórias e antitumorais, mas ainda pouco estudado em modelos animais. Objetivos: Avaliar os efeitos da administração do extrato alcoólico de G. brasiliensis em modelo de carcinogênese induzida por benzopireno. Material e Método: O extrato bruto foi obtido por percolação com o uso de 20 g das folhas secas e trituradas de G. brasiliensis e 100 ml de etanol a 70%, por 24h. Ratos Wistar foram divididos em 3 grupos, um controle sem indução ou tratamento, um induzido pelo benzopireno (100 mg/kg, diluído em DMSO e administrado intraperitonealmente, uma única aplicação) e um grupo tratado por gavagem (1 ml) com extrato de bacupari a 4% (3x/semana, por 7 semanas, a partir da 15o semana da indução. Os animais de todos os grupos foram eutanasiados após 21 semanas para coleta dos pulmões que foram processados para análises histopatológicas (HE) e histoquímicas (Azul de Toluidina e Azul de Alcian Safranina) para quantificação dos mastócitos. Resultados: Os resultados das análises histopatológicas mostraram desorganização do parênquima pulmonar, aumento de tecido linfático associado aos brônquios, grande influxo de células inflamatórias e regiões de displasia. Pela coloração de azul de toluidina os mastócitos foram identificados na forma intacta e desgranulada (em processo de ativação). Na coloração conjunta Azul de Alcian e Safranina, os mastócitos corados em azul representam as fases iniciais do processo de maturação, enquanto os corados em vermelho estão maduros. A quantificações evidenciaram maior quantidade de mastócitos desgranulados, azul de Alcian e mistos (corados pelo Azul de Alcian e Safranina) nos grupos tratados com o extrato. Conclusão: Os dados indicam que o ambiente tumoral nos animais tratados mostra maior modulação dos mastócitos, com mais células jovens e ativadas. Mais análises serão realizadas para verificar se a ativação dos mastócitos promovida pelo tratamento com o extrato ocorre para contenção ou promoção do processo tumoral.(AU)
Introduction: Benzopyrene is one of the main polycyclic aromatic hydrocarbons present in the environment and with a high carcinogenic capacity, being, therefore, used in in vivo models of lung carcinogenesis. Investigations have shown the involvement of mast cells in modulating the tumor environment and that anti-inflammatory drugs can reduce the incidence of lung cancer. Among the therapeutic possibilities are herbal medicines. Garcinia brasiliensis extract, popularly known as bacupari, shows anti-inflammatory and antitumor properties, but still little studied in animal models. Objectives: To evaluate the effects of the administration of the alcoholic extract of G. brasiliensis in a model of carcinogenesis induced by benzopyrene. Material and Methods: The crude extract was obtained by percolation using 20 g of dried and crushed leaves of G. brasiliensis and 100 ml of 70% ethanol for 24 hours. Wistar rats were divided into 3 groups, a control without induction or treatment, one induced by benzopyrene (100 mg/kg, diluted in DMSO and administered intraperitoneally, a single application) and a group treated by gavage (1 ml) with bacupari extract. at 4%...(AU)
Introducción: El benzopireno es uno de los principales hidrocarburos aromáticos policíclicos presentes en el ambiente y tiene una elevada capacidad carcinogénica, por lo que se utiliza en modelos in vivo de carcinogénesis pulmonar. Las investigaciones han demostrado la implicación de los mastocitos en la modulación del entorno tumoral y que los fármacos antiinflamatorios pueden reducir la incidencia del cáncer de pulmón. Entre las posibilidades terapéuticas están las fitoterapias. El extracto de Garcinia brasiliensis, conocido popularmente como bacupari, muestra propiedades antiinflamatorias y antitumorales, pero todavía está sido poco estudiado en modelos animales. Objetivos: Evaluar los efectos de la administración del extracto alcohólico de G. brasiliensis en el modelo de carcinogénesis inducido por el benzopireno. Material y métodos: El extracto crudo se obtuvo por percolación utilizando 20 g de hojas de G. brasiliensis secas y trituradas y 100 ml de etanol al 70%, durante 24h. Las ratas Wistar fueran divididas en 3 grupos, uno de control sin inducción ni tratamiento, otro inducido por benzopireno (100 mg/kg, diluido en DMSO y administrado por vía intraperitoneal, una sola aplicación) y un grupo tratado por gavage (1ml) con extracto de bacupari 4% (3x/semana, durante 7 semanas, desde la 15ª semana de inducción). Los animales de todos los grupos fueron eutanasiados después de 21 semanas para recoger los pulmones que se procesaron para los análisis histopatológicos (HE) e histoquímicos (azul de toluidina y azul de safranina) para la cuantificación de los mastocitos. Resultados: Los resultados de los análisis histopatológicos mostraron desorganización del parénquima pulmonar, aumento del tejido linfoide asociado a los bronquios, gran afluencia de células inflamatorias y regiones de displasia. Mediante la tinción con azul de toluidina, los mastocitos se identificaron como intactos y degranulados (en proceso de activación). En la tinción conjunta de azul Alcian y Safranina, los mastocitos teñidos de azul representan las fases iniciales del proceso de maduración, mientras que los teñidos de rojo son maduros. La cuantificación mostró una mayor cantidad de mastocitos degranulados, azul Alcian y mixtos (teñidos con azul Alcian-Safranin) en los grupos tratados con el extracto. Conclusión: Los datos indican que el entorno del tumor en los animales tratados muestra una mayor modulación de los mastocitos, con más células jóvenes y activadas. Se llevarán a cabo más análisis para verificar si la activación de los mastocitos promovida por el tratamiento con el extracto se produce para contener o promover el proceso tumoral.(AU)
Assuntos
Animais , Plantas Medicinais , Carcinogênese , Neoplasias Pulmonares , Benzopirenos/administração & dosagem , Ratos Wistar , Anti-InflamatóriosRESUMO
Benzopyrene is one of the main polycyclic aromatic hydrocarbons with carcinogenic capacity. Research has shown that anti-inflammatory drugs can reduce the incidence of lung cancer. In this scenario, we highlight piperlongumin (PL), an alkaloid from Piper longum with anti-inflammatory properties. Therefore, our aim was to study the effect of PL administration in a model of pulmonary carcinogenesis induced by benzopyrene in Balb/c mice. Animals were divided into 3 groups (n = 10/group): sham (10% DMSO), induced by benzopyrene (100 mg/kg, diluted in DMSO) without treatment (BaP) for 12 weeks and induced by benzopyrene and treated with PL (BaP/PL) (2 mg/kg in 10% DMSO) from the eighth week post-induction. Animals were weighed daily and pletsmography was performed in the 12th week. Genotoxicity and hemoglobin levels were analyzed in blood and quantification of leukocytes in bronchoalveolar lavage (BAL). Lungs were collected for histopathological evaluation, immunohistochemical studies of annexin A1 (AnxA1), cyclooxygenase 2 (COX-2), anti-apoptotic protein Bcl-2 and nuclear transcription factor (NF-kB) and also the measurement of interleukin cytokines (IL)-1ß, IL-17 and tumor necrosis factor (TNF) -α. Treatment with PL reduced the pulmonary parameters (p < 0,001) of frequency, volume and pulmonary ventilation, decreased lymphocytes, monocytes and neutrophils in BAL (p < 0,05) as well as blood hemoglobin levels (p < 0,01). PL administration also reduced DNA damage and preserved the pulmonary architecture compared to the BaP group. Moreover, the anti-inflammatory effect of PL was evidenced by the maintenance of AnxA1 levels, reduction of COX-2 (p < 0,05), Bcl-2 (p < 0,01) and NF-kB (p < 0,001) expressions and decreased IL-1ß, IL-17 (p < 0,01) and TNF-α (p < 0,05) levels. The results show the therapeutic potential of PL in the treatment of pulmonary anti-inflammatory and anti-tumor diseases with promising therapeutic implications.
Assuntos
Anti-Inflamatórios/farmacologia , Animais , Anexina A1/metabolismo , Benzo(a)pireno/metabolismo , Benzopirenos , Líquido da Lavagem Broncoalveolar , Carcinógenos/metabolismo , Ciclo-Oxigenase 2/metabolismo , Citocinas/metabolismo , Inflamação/metabolismo , Interleucina-1beta , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Neutrófilos/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Astrocytomas represent the majority of cerebral gliomas. Studies show that the anti-inflammatory protein Annexin-A1 (ANXA1) is associated with the tumor invasion process and that its actions can be mediated by the receptor for formylated peptides (FPR). Therefore, we evaluated the expression of ANXA1, the receptor FPR2 and matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) in brain astrocytomas. Detection of proteins was performed in sections of diffuse astrocytomas (grade II), anaplastic astrocytomas (grade III) and glioblastomas (GBM, grade IV) and quantifications were made by densitometry. Our analyses showed increased expression of ANXA1 in astrocytomas of all grades, but especially in GBM. The expression of FPR2 is similar to that found for ANXA1, being higher in GBM. Immunostaining for MMPs is also stronger as the degree of malignancy increases, especially with respect to MMP-9. The positive correlation between ANXA1/FPR2 and ANXA1/MMP-9 was observed in all tumors studied. The data indicate the possible action of ANXA1 and FPR2 on the development and progression of astrocytomas, related to increased expression of MMP-9. Thereby, ANXA1 and FPR2 are involved in the biology and malignancy of diffuse astrocytic tumors.
Assuntos
Anexina A1/biossíntese , Astrocitoma/patologia , Biomarcadores Tumorais/biossíntese , Neoplasias Encefálicas/patologia , Receptores de Formil Peptídeo/biossíntese , Receptores de Lipoxinas/biossíntese , Adulto , Idoso , Feminino , Humanos , Masculino , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Pessoa de Meia-IdadeRESUMO
Mast cells (MCs) participate in all stages of skin healing and one of their mediators is the Annexin A1 protein (AnxA1), linked to inflammation, proliferation, migration and apoptosis processes, but not studied in thermal burns yet. Therefore, our objectives were to evaluate the behavior of MCs and AnxA1 in a second degree burn model, treated or not with silver sulfadiazine 1% (SDP 1%) and associated to macrophages quantification and cytokines dosages. MCs counts showed few cells in the early stages of repair but increased MCs in the final phases in the untreated group. The normal skin presented numerous tryptase-positive MCs that were reduced after burning in all analyzed periods. Differently, few chymase-positive MCs were observed in the early stages of healing, however, increased chymase-positive MCs were found at the final phase in the untreated group. MCs also showed high immunoreactivity for AnxA1 on day 3 in both groups. In the tissue there was a strong protein expression in the early stages of healing, but in the final phases only in the SDP treated animals. TNF-α, IL-1ß, IL-6, IL-10 and MCP-1 levels and macrophages quantification were increased in inflammation and reepithelialization phases. Reduced IL-1ß, IL-6 and IL-10 levels and numerous macrophages occurred in the treated animals during tissue repair. Our results indicate modulation in the profile of MCs and AnxA1expression during healing by the treatment with SDP 1%, pointing them as targets for therapeutic interventions on skin burns.