Perfis fitoquímicos, microbiológicos, citotóxicos e antiinflamatórios do extrato alcoólico de folhas de atemoia (annona squamosa, l. X annona cherimola) / Phytochemical, microbiological, cytotoxic and anti-inflammatory profiles of the alcoholic extract of atemoy leaves (annona squamosa, l. X annona cheremola) / Perfiles fitoquímico, microbiológico, citotóxico y antiinflamatorio del extracto alcohólico de hojas de atemoy (annona squamosa, l. X annona cheremola)

Introdução: Os extratos alcoólicos de plantas com potencial medicinal possuem compostos com alto potencial antiinflamatório e aplicação terapêutica. Dentre as espécies vegetais de interesse destacamos a atemoia, um hibrido de Annona cherimola, Mill e Annonas quamosa, L. Objetivo: Caracterizar o extrato alcoólico bruto de folhas da atemoia em ensaios fitoquímicos, microbiológicos, citotóxicos e no modelo de peritonite. Método: O extrato de folhas de atemoia foi obtido por percolação, com o uso de 20g de folhas secas e trituradas com 100 ml de álcool de cereais. Na padronização dos extratos foram utilizadas análises de idenficação de componentes químicos, os taninos, flavonoides, saponinas e alcaloides, por meio de difrentes reações químicas e os antioxidantes foram analisados pela atividade do DPPH. Diferentes cepas bacterianas foram avaliadas quanto a sensililidade ao extrato em várias concentrações. A citotoxicidade das diferentes concentrações do extrato também foi analisada por hemólise e pelo teste da membrana corioalantide (CAM). Ainda, a capacidade anti-inflamatória do extrato a 4% foi avaliada na peritonite induzida por endotoxina em ratas. Resultados: As análises fitoquímicas indicaram a presença de alcaloides, taninos e flavonoides e ausência de saponinas além da alta capacidade antioxidante. Na concentração de 100% o extrato inibiu a proliferação de bactérias Gram-positivas e Gram-negativas. Os testes de hemólise e CAM mostraram ausência de citotoxicidade na concentração de 4% e elevada citotoxicidade a partir de 10%. No modelo de peritonite, a administração do extrato na concentração de 4% inibiu a migração de neutrófilos para a cavidade peritoneal. Conclusão: O extrato alcoólico das folhas da atemoia mostra importante perfil anti-inflamatório e antioxidante o que estimula estudos futuros para sua aplicação farmacológica. (AU)

Introduction: The alcohol extracts of plants with medicinal potential have compounds with high anti-inflammatory potential and therapeutic application. Among the plant species of interest, we highlight athemoia, a hybrid of Annona cherimola, Mill and Annonas quamosa, L. Objective: To characterize the raw alcoholic extract of athemoious leaves in phytochemical, microbiological, cytotoxic tests and in the peritonitis model. Method: The extract of atemoia leaves was obtained by percolation, with the use of 20g of dried leaves and crushed with 100 ml of cereal alcohol. In the standardization of the extracts were used idenfication analyses of chemical components, tannins, flavonoids, saponins and alkaloids, by means of diffractive chemical reactions and antioxidants were analyzed by DPPH activity. Different bacterial strains were evaluated regarding the sensitivity to the extract in different concentrations. Cytotoxicity of the different concentrations of the extract were also analyzed by hemolysis and by the corioalantide membrane (CAM) test and the anti-inflammatory capacity of the 4% extract was evaluated in endotoxin-induced peritonitis in rats. Results: Phytochemical analysis indicated the presence of alkaloids, tannins and flavonoids and absence of saponins and high antioxidant capacity. At 100% concentration the extract inhibited the proliferation of gram-positive and gram-negative bacteria. Hemolysis and CAM tests showed no cytotoxicity at 4% concentration and high cytotoxicity at 10% level. In the peritonitis model, administration of the extract at the 4% concentration inhibited the migration of neutrophils to the peritoneal cavity. Conclusion: The alcoholic extract of the leaves of athemoia shows an important anti-inflammatory and antioxidant profile which stimulates future studies for its pharmacological application.(AU)

Introducción: Los extractos alcohólicos de plantas con potencial medicinal presentan compuestos con alto potencial antiinflamatorio y aplicación terapéutica. Entre las especies vegetales de interés destacamos la atemoya, un híbrido de Annona cherimola, Mill y Annonas quamosa L. Objetivo: Caracterizar el extracto alcohólico crudo de hojas de atemoya en ensayos modelo fitoquímicos, microbiológicos, citotóxicos y de peritonitis. Método: El extracto de hojas de atemoya se obtuvo por percolación, utilizando 20g de hojas secas y trituradas con 100 ml de alcohol de grano. En la estandarización de los extractos se utilizaron análisis para identificar componentes químicos, taninos, flavonoides, saponinas y alcaloides, a través de diferentes reacciones químicas y los antioxidantes fueron analizados por la actividad de DPPH. Se evaluó la sensibilidad al extracto de diferentes cepas bacterianas a diferentes concentraciones. También se analizó la citotoxicidad de las diferentes concentraciones del extracto por hemólisis y por el test de membrana de corioalantida (CAM) y se evaluó la capacidad antiinflamatoria del extracto al 4% en peritonitis inducida por endotoxinas en ratas. Resultados: Los análisis fitoquímicos indicaron la presencia de alcaloides, taninos y flavonoides y la ausencia de saponinas y alta capacidad antioxidante. A una concentración del 100%, el extracto inhibió la proliferación de bacterias Gram-positivas y Gram-negativas. Las pruebas de hemólisis y CAM no mostraron citotoxicidad a una concentración del 4% y alta citotoxicidad a partir del 10%. En el modelo de peritonitis, la administración del extracto a una concentración del 4% inhibió la migración de neutrófilos a la cavidad peritoneal. Conclusión: El extracto alcohólico de hojas de atemoya muestra un importante perfil antiinflamatorio y antioxidante, lo que estimula futuros estudios para su aplicación farmacológica.(AU)

Modulation of the endogenous Annexin A1 in a cigarette smoke cessation model: Potential therapeutic target in reversing the damage caused by smoking?

BACKGROUND: Smoking cessation may help in the reversal of inflammation and damage caused by smoking. The endogenous annexin A1 (AnxA1) protein has anti-inflammatory effects which instigates the understanding of its role in the attenuation of inflammatory processes caused by smoking. MATERIAL AND METHODS: Wistar rats were exposed to cigarette smoke for 8 weeks. After the exposure period, one of the groups remained other 8 weeks in the absence of smoke. Animals not exposed to smoke were used as control. Blood, trachea and lungs were obtained for histopathological, immunohistochemical and biochemical analyses. RESULTS: Loss of cilia of the tracheal lining epithelium was found by smoke exposure, but smoking cessation led to recovery of the tracheal epithelium. Similarly, chronically exposed-to-smoke animals showed increased lymphocytes and macrophages in bronchoalveolar lavage and higher levels of glucose and gamma-GT in their blood. Reduction of lymphocytes, glucose and gamma-GT occurred after smoking cessation. In addition, IL-1ß, IL-6, IL-10, TNF-α and MCP-1 levels were elevated by smoke exposure. Smoking cessation significantly reduced the levels of IL-1ß, IL-6 and MCP-1 but increased the IL-10 concentration. Numerous mast cells and macrophages were observed in the lung of chronically exposed-to-smoke animals with reduction by smoking cigarette abstinence. AnxA1 increased expression and concomitant NF-κB reduction were found in the smoking cessation group. CONCLUSION: Our results showed that cigarette abstinence promoted partial recovery of the inflammatory process. The attenuation of the inflammatory profile may be associated with the overexpression of AnxA1 protein.

Heterogeneity of mast cells and expression of Annexin A1 protein in a second degree burn model with silver sulfadiazine treatment.

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.

Análise do perfil dos mastócitos em fibroadenomas e carcinomas ductais de mama / Análisis del perfil de los mastócitos en el fibroadenoma de mama y carcinomas ductales / Mast cells profile analysis in fibroadenomas and ductal breast carcinomas

Introdução: O câncer de mama acomete milhares de mulheres no Brasil e no mundo, o que estimula pesquisas nesse campo. Osmastócitos (MCs) são células infl amatórias relacionadas ao microambiente tumoral e que podem variar de acordo com o tipo e oestadiamento dos tumores. Objetivo: Por essas razões, buscou-se analisar o comportamento dos MCs em biópsias de tumores de mamabenignos, os fi broadenomas, e malignos, os carcinomas de graus 1, 2 e 3 metastáticos ou não. Material e Método: Estudo realizadopor meio da quantifi cação dessas células, avaliação do seu estado de ativação, após coloração com Azul de Toluidina bem como, daheterogeneidade, pela imuno-histoquímica para as proteases triptase e quimase. Resultados: Modulação no número de MCs e do estadode ativação nos fi broadenomas e carcinomas de graus 1, 2 e 3, com e sem metástases. Conclusão: Evidenciou-se a participação ativadessas células no desenvolvimento tumoral.

Introduction: Breast cancer affects thousands of women in Brazil and over around the world, and this fact stimulates research in this fi eld.Mast cells (MCs) are infl ammatory cells related to the tumoral microenvironment and they may vary according to the type and staging oftumors. Objective: For these reasons, we looked for analyzing the profi le of MCs in biopsies of benign breast tumors, the fi broadenomas,as well as of malignant ones, the carcinomas of 1, 2 and 3 degree, metastatic or not. Materials and method: The study was carriedout by quantifying these cells, evaluating their activation state, after staining with Toluidine Blue, as well as their heterogeneity, byimmunohistochemistry for the proteases tryptase and kinase. Results: Modulation in the MCs number and activation state in bothfi broadenomas and carcinomas of 1, 2 and 3degree, with and without metastases. Conclusion: we demonstrated the active participationof these cells in tumor development.

Introducción: El cáncer de mama afecta a miles de mujeres en Brasil y en el mundo, lo que estimula las investigaciones en este campo.Los mastocitos (MCs) son células infl amatorias relacionadas con el microambiente tumoral y que pueden variar de acuerdo con el tipoy la estadifi cación de los tumores. Objetivo: Por estas razones, se buscó analizar el comportamiento de los MCs en biopsias de tumoresde mama benignos, los fi broadenomas, y los malignos, los carcinomas de grados 1, 2 y 3 metastáticos o no. Material y Método: Estudiorealizado por medio de la cuantifi cación de esas células, evaluación de su estado de activación, después de la coloración con Azul deToluidina así como de la heterogeneidad, por la inmuno-histoquímica para las proteasas, triptasa y quimase. Resultados: Modulación enel número de MCs y del estado de activación en los fi broadenomas y carcinomas de grados 1, 2 y 3, con y sin metástasis. Conclusión: Seevidenció la participación activa de estas células en el desarrollo tumoral.

Anexina a1 e metaloproteinases de matriz em neoplasias uterinas / Anexina a1 y metaloproteinasas de matriz en neoplasias uterinas / Annexin a1 and matrix metalloproteinases in uterine neoplasias

Introdução: As neoplasias uterinas são causas importantes de desconforto, infertilidade e óbito entre as mulheres no Brasil e no mundo.Nesse cenário, destaca-se a proteína anti-inflamatória anexina A1 (ANXA1) que tem sido associada com a progressão em tumoresindicando que essa proteína regula o processo de migração/invasão celular. Objetivo: Analisar, por imuno-histoquímica, a expressãoda ANXA1 e sua correlação com as metaloproteinases de matriz (MMP-2 e MMP-9), bem como com o fator de crescimento vascularendotelial (VEGF) no leiomioma e no adenocarcinoma de endométrio. Material e Método: As análises foram realizadas em biópsias deleiomiomas e adenocarcinomas. Amostras de útero normal foram usadas como controle. A quantificação da expressão das proteínasfoi feita por densitometria. Resultados: Nossos resultados mostraram aumento da expressão da proteína nos tumores benignos emalignos. Conclusão: Similarmente, observamos expressões aumentadas de MMPs e VEGF nas neoplasias uterinas, o que parece indicara participação da proteína ANXA1 nos processos de proliferação e invasão celular.

Introduction: Uterine neoplasias are major causes of discomfort, infertility and death among women in Brazil and worldwide. In thisscenario, we highlight the anti-inflammatory protein annexin A1 (ANXA1) that has been associated with progression in tumors suggestingthat this protein regulates the process of cell migration/invasion. For these reasons. Objective: To analyze, by immunohistochemistry,the expression of ANXA1 and its correlation with matrix metalloproteinases (MMP-2 and MMP-9), as well as with the vascular endothelialgrowth factor (VEGF) in leiomyoma and endometrial adenocarcinoma. The analyzes were performed on biopsies from leiomyomasand adenocarcinomas. Normal uterus samples were used as controls. The quantification of the protein expression was performed bydensitometry. Results: Our results showed increased protein expression in both benign and malignant tumors. Conclusion: Similarly, weobserved increased expression of both MMPs and VEGF in uterine neoplasias which seems to indicate the participation of ANXA1 proteinin cell proliferation and invasion processes.

Introducción: El cáncer uterino son las principales causas de malestar, la infertilidad y la muerte entre las mujeres en Brasil y en todo elmundo. En este escenario, la proteína anti-inflamatoria anexina A1 (ANXA1) que se ha asociado con la progresión de los tumores, lo queindican que esta proteína regula la invasión migración / célula. Objetivo: analizar, mediante inmunohistoquímica, la expresión de ANXA1y su correlación con las metaloproteinasas de matriz (MMP-2 y MMP-9), así como el factor de crecimiento endotelial vascular (VEGF) enleiomioma y adenocarcinoma endometrial. Material y Método: Los análisis se realizaron en biopsias de leiomiomas y adenocarcinomas.Muestras de útero normal se utilizaron como controles. La cuantificación de la expresión de la proteína se realizó por densitometría.Resultados: Nuestros resultados mostraron un aumento de expresión de la proteína en los tumores benignos y malignos. Conclusión: Delmismo modo, se observó aumento de la expresión de MMPs y el cáncer uterino VEGF, lo que parece indicar la participación de la proteínaANXA1 en la proliferación y la invasión celular.