Effects of elderflower extract enriched with polyphenols on antioxidant defense of salmon leukocytes

BACKGROUND: In fish farming, the plant extracts containing antioxidant compounds have been added to the diet for enhancing pathogen resistance. In vitro studies evaluating the antioxidant effect of herbal extracts on fish cell models have focused on ROS production and the respiratory burst mechanism. However, the effects on enzymatic antioxidant defense on salmon leukocytes have not been evaluated. This study aims to evaluate the enzymatic antioxidant defense and ROS-induced cell damage in Salmon Head Kidney-1 (SHK-1) cell line exposed to polyphenol-enriched extract from Sambucus nigra flowers. RESULTS: Firstly, the Total Reactive Antioxidant Power (TRAP) assay of elderflower polyphenol (EP) was evaluated, showing 459 and 489 times more active than gallic acid and butyl hydroxy toluene (BHT), respectively. The toxic effect of EP on salmon cells was not significant at concentrations below 120 mg/ mL and no hemolysis activity was observed between 20 and 400 mg/mL. The treatment of SHK-1 cell line with EP decreased both the lipid peroxidation and protein oxidation induced by H2O2, which could be associated with decreasing oxidative stress in the SHK-1 cells since the GSH/GSSG ratio increased when only EP was added. CONCLUSIONS: These results suggest that plant extracts enriched with polyphenols could improve the enzymatic antioxidant defense of salmon leukocytes and protect the cells against ROS-induced cell damage

Chemical properties and assessment of the antioxidant capacity of native species from the genus Ugni / Características químicas y evaluación de la capacidad antioxidante de especies del género Ugni

Introduction: Ugni molinae Turcz., Ugni candollei Barn. (Berg) and Ugni selkirkii (Hook. et Arn.) Berg are Chilean species that share morphological characteristics and they are distributed in geographic locations with very diverse habitats. Its is considered important for the characterization of the Chilean flora to determine if there are similarities in the sort of chemical compounds among species with close morphological relations, growing in different habitats and their consequent biological activity. Objective: to assess the chemical composition and the antioxidant capacity of leaf extracts from the Chilean species of the genus Ugni, and to compare with the U. molinae characteristics. Methods: composition of chemical compounds was determined by chromatographic methods (HPLC-ESI-MS). The antioxidant capacity was assessed by DPPH, ABTS and by stabilization of the hydroxyl radical. Results: as expected, given the great morphological similarity existing among the three species of Ugni that grow in Chile, similarities were found in their chemical composition. Nevertheless, it was also expectable to find variations among them. Thus, U. candollei and U. selkirkii are the species that present greater content and variety of phenolic and terpenic compounds. These species exert greater antioxidant capacity in comparison toU. molinae. U. candollei hightlights for its flavonoid content such as glycosides and quercetin derivatives, and the species U. selkirkii, is important in galotannins. U. molinae is characterized in ellagic acids derivatives. Conclusion: these data and the morphological characteristics could become a useful toll in order to determine the closeness degree among these species(AU

Introducción: Ugni molinae Turcz., Ugni candollei (Barn.) Berg y Ugni selkirkii (Hook. et Arn.) Berg son especies chilenas que comparten características morfológicas y se distribuyen en lugares geográficos con muy diversos hábitats. Se considera importante para la caracterización de la flora chilena determinar si hay similitudes en el tipo de compuestos químicos entre especies con relaciones morfológicas cercanas, que crecen en diferentes hábitats y su actividad biológica consecuente. Objetivo: evaluar la composición química y la capacidad antioxidante de extractos de hojas de las especies chilenas del género Ugni, y compararlas con las características de la especie U. molinae. Métodos: la composición química se determinó por métodos cromatográficos (HPLC-ESI-MS). La capacidad antioxidante se evaluó por los métodos DPPH, ABTS, y la estabilización del radical hidroxilo. Resultados: como era de esperar, dada la gran similitud morfológica existente entre las tres especies de Ugni que crecen en Chile, se encontraron similitudes en su composición química. Sin embargo, también era esperable encontrar variaciones entre ellas. Por lo tanto, U. candollei y U. selkirkii son las especies que presentan mayor contenido y variedad de compuestos fenólicos y terpénicos. Estas especies ejercen una mayor capacidad antioxidante en comparación con U. molinae. U. candollei se caracteriza por su contenido en flavonoides como quercetina glucósidos y sus derivados, y la especie U. selkirkii, por la presencia de galotaninos. U. molinae se caracteriza por contener derivados del ácido elágico. Conclusión: estos datos y las características morfológicas podrían convertirse en una herramienta útil para determinar el grado cercanía entre estas especies(AU)

An HPLC-UV and HPLC-ESI-MS based method for identification of anti-inflammatory triterpenoids from the extracts of Ugni molinae / Método CLAE-UV y CLAE-IES-MS para la identificación de triterpenoides anti-inflamatorios de los extractos de Ugni molinae

The aim of this study was to establish an analytical method to detect the presence of the responsible triterpenoids of the anti-inflammatory activity of the leaves of Ugni molinae (murtilla). Successive leaves extracts of EtOAc (EAE) and ethanol (TEE) were prepared, obtaining for the first time from TEE a triterpenoid-rich sub-fraction (TF). The topical anti-inflammatory activity of TF was assessed (43.3 percent at 1 mg/ear) by means of the TPA-induced mouse ear oedema model, which was compared to EAE (83.1 +/- 3.2 percent) and TEE (78.3 +/- 11.8 percent) activities, both previously evaluated by us. These extracts were characterized in their triterpenoids by HPLC-UV and HPLC-ESI-MS. We demonstrated that TF has triterpenoids responsible in part of the anti-inflammatory activity, among them, madecassic and maslinic acids. These two compounds have been reported for the first time for this species. ED50 for madecassic and alphitolic acids are also here reported.

El objetivo de este trabajo fue establecer un método analítico para determinar la presencia de los triterpenoides responsables de la actividad anti-inflamatoria de las hojas de Ugni molinae (murtilla). Fueron preparados los extractos seriados de EtOAc (EAE) y etanólico (TEE) desde sus hojas, obteniendo desde el TEE por primera vez una sub-fracción rica en triterpenoides (TF). Se demostró la actividad anti-inflamatoria tópica del TF por el modelo de edema de oreja de ratón inducida por TPA (43,3 por ciento a 1 mg/oreja), la cual fue comparada con las de los EAE (83,1 +/- 3,2 por ciento) y TEE (78,3 +/- 11,8 por ciento) determinadas en nuestros estudios previos. Dichos extractos fueron caracterizados en sus triterpenoides por CLAE-UV y CLAE-IES-MS. Demostramos que el TF contiene triterpenoides responsables en parte de la actividad anti-inflamatoria, entre ellos, los ácidos madecásico y maslínico, reportados por primera vez para esta especie. Se informan además las DE50 para los ácidos madecásico y alfitólico.

Microtubule proteins and their post-translational forms in the cerebrospinal fluid of patients with paraparesis associated with HTLV-I infection and in SH-SY5Y cells: An in vitro model of HTLV-I-induced disease

HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is characterized by axonal degeneration of the corticospinal tracts. The specific requirements for transport of proteins and organelles to the distal part of the long axon are crucial in the corticospinal tracts. Microtubule dysfunction could beinvolved in this disease, configuring an axonal transport disease. We measured tubulin and its posttranslational modified forms (acetylated and tyrosinated) in CSF of patients and controls, as well as tau and its phosphorylated forms. There were no significant differences in the contents of tubulin and acetyl-tubulinbetween patients and controls; tyrosyl-tubulin was not detected. In HAM/TSP, tau levels were significantly reduced, while the ratio of pT181/total tau was higher in patients than in controls, this being completely different from what is reported in other neurodegenerative diseases. Phosphorylation at T181 was also confirmed by Mass Spectrometry analysis. Western Blotting with monospecific polyclonal antibodies against pS199, pT205, pT231, pS262, pS356, pS396, pS404 and pS422 did not show differences in phosphorylation in these residues between patients and controls. Treating human SH-SY5Y neuroblastoma cells, a well-known in vitro neurite retraction model, with culture supernatant of MT-2 cells (HTLV-I infected cell line that secretes theviral Tax protein) we observed neurite retraction and an increase in tau phosphorylation at T181. A disruptionof normal phosphorylation of tau protein in T181 could result in its dysfunction, contributing to axonal damage.