Protein from Hevea brasiliensis "Hev b 13" latex attenuates systemic inflammatory response and lung lesions in rats with sepsis / Proteína do latex de Hevea brasiliensis "Hev b 13" atenua resposta inflamatória sistêmica e lesão pulmonar de ratos com sepse

Abstract Sepsis induces a severe systemic inflammatory response that may result in multiple organ dysfunction and death. Studies using a protein derived from natural Hevea brasiliensis (rubber tree) latex, denominated Hev b 13, have demonstrated important anti-inflammatory effects, but no data have been published regarding its effects on sepsis. The aim of this study was to investigate the effects of Hev b 13 on the inflammatory response and lung lesions of septal rats. Male Wistar rats were submitted to cecal ligation and puncture (CLP), randomized into groups and treated with subcutaneously administered doses of 0.5/2.0/3.0 mg/Kg of Hev b 13. Next, animals were subdivided into three different points in time (1, 6 and 24 hours after treatments) for collection of blood samples and euthanasia accompanied by organ removal. Total and differential leukocyte counts, cytokine dosage and histological assessment were analyzed. Treatment with Hev b 13 resulted in a significant decline in total and differential leukocytes as well as suppression of TNF-α and IL-6 production, associated with the increase in IL-10 and IL-4 in plasma and lung tissue. Moreover, it reduced morphological and pathological changes found in the lungs, including neutrophil infiltration, edema and alveolar thickening. The present study concluded that Hev b 13 exerts anti-inflammatory effects and attenuates lung lesions in septal rats, showing potential for clinical application.

Resumo Sepse induz uma resposta inflamatória sistêmica grave podendo resultar em disfunção de múltiplos órgãos e morte. Pesquisas utilizando uma proteína derivada do látex natural de Hevea brasiliensis (seringueira), denominada Hev b 13 tem demonstrado importantes efeitos anti-inflamatórios, mas nenhum dado foi publicado dos seus efeitos na sepse. O objetivo deste estudo foi investigar os efeitos da Hev b 13 na resposta inflamatória e na lesão pulmonar de ratos com sepse. Ratos machos da linhagem Wistar foram submetidos a ligação e perfuração do ceco (LPC), randomizados em grupos e tratados com as doses 0,5/2,0/3,0 mg/Kg de Hev b 13 subcutâneo. Após subdividiu-se os animais em três pontos diferentes de tempo (1, 6 e 24 horas após os tratamentos) para coleta de amostras sanguíneas e eutanásia com remoção dos órgãos. Contagem total e diferencial de leucócitos, dosagem de citocinas e avaliação histológica foram analisadas. O tratamento com a Hev b 13 resultou em diminuição significativa de leucócitos totais e diferenciais bem como suprimiu a produção de TNF-α e IL-6, associado ao aumento de IL-10 e IL-4 no plasma e tecido pulmonar. Além disso, reduziu as alterações morfológicas e patológicas encontradas nos pulmões, incluindo infiltrado de neutrófilos, edema e espessamento alveolar. Este estudo concluiu que a Hev b 13 tem efeitos anti-inflamatórios e atenua lesões pulmonares em ratos com sepse, apresentando potencialidades para aplicabilidade clínica.

Efecto de la temperatura de conservación sobre la expresión de proteinas en granos de polen de chenopodium album l / Effect of conservation temperature on the expression of proteins in pollen grains of chenopodium album l

Los componentes activos del polen a partir del cual se obtienen los extractos alergénicos pueden variar considerablemente deacuerdo al momento, el lugar dónde se recolecta y el períodocomprendido entre la recolección y su utilización.El presente trabajo pretende evaluar si la temperatura de conservacióndel grano de polen influye en la expresión de proteínasy en la antigenicidad de las mismas, al momento de prepararun extracto alergénico. La especie elegida para estudio fue Chenopodiumalbum L. ya que es de gran interés alergológico en laciudad de Bahía Blanca. Los granos de polen se conservaron a temperatura ambiente, 4 °Cy -18 °C por el término de dos meses. El contenido proteico de losextractos se determinó por el Método de Bradford. La expresiónproteica y la antigenicidad se estudiaron mediante electroforesisvertical Tricina-PAGE-SDS 12, 5 % e Inmunoblot respectivamente.Los resultados obtenidos demuestran que la concentración proteicatotal fue menor para los extractos obtenidos del polen conservadoa temperatura ambiente que para las otras dos condiciones.La expresión de proteínas varía cuantitativamente en todoslos extractos y si bien la expresión cualitativa prácticamente seconserva, aparece para el polen conservado a temperatura ambiente,una banda de PM menor a 12 kDa. Esta banda podríaser consecuencia de la degradación proteica que experimenta elpolen a esa temperatura de almacenamiento. En cuanto a la antigenicidadde los extractos no hay diferencias cualitativas aunquepueden apreciarse diferencias cuantitativas significativas.Concluimos que la conservación del polen a 4°C o a -18°C seríanlas más adecuadas, ya que permiten obtener una mayor concentraciónproteica partiendo de la misma masa de polen.

The active components from which pollen allergen extracts are obtainedcan change considerably according to the time or the placewhere collect and the time period between harvesting and use.This work aims to assess if the storage temperature of the pollengrain influences protein expression and its antigenicity when preparingan allergen extract. The species chosen for our study wasChenopodium album L, since it is of great allergologic interest inthe city of Bahía Blanca.Pollen grains were stored at room temperature 4ºC and -18ºC ,for a period of two months. The protein content of the extractswas determined by the Bradford method. Protein expressionand antigenicity were studied by vertical electrophoresis TricineSDS-PAGE 12, 5% and Immunoblot. The obtained results show that the total protein concentrationwas lower in the extracts of pollen stored at room temperaturethan in those under two conditions. Protein expression differsquantitatively in all extracts and even if the qualitative expressionis kept practically the same, in the Tricine SDS-PAGE geland the pollen stored at room temperature there appear a bandof MW inferior to 12 kDa. This band could result from the proteindegradation experienced by pollen stored at that temperature.As regards extract antigenicity, there are no qualitative differenceseven though there are significant quantitative differences.We conclude that pollen preservation at 4ºC or -18ºC would bethe most appropriate since it allows greater protein concentrationfor the same mass of pollen.