Actividad antiinflamatoria de un extracto polifenólico de hueso de olivas en la línea celular de monocitos humanos THP1-XBLUE-CD14

Abstract

El objetivo de este trabajo es evaluar la actividad antiinflamatoria de un extracto de naturaleza polifenólica de huesos de oliva. Material y métodos: Se incubó la línea celular THP1-XBlue-CD14 (invivogen), 80.000 células/pocillo, provocando inflamación (activación de NF-kb) mediante 0.1 μg/ml LPS (lipopolisacárido de E. coli) durante 24 horas. Se evaluó la presencia del extracto (10 y 50 mg/l, concentraciones bioseguras) durante 2 horas a 37 ºC, previa (efecto preventivo) y posterior a la activación proinflamatoria (efecto terapéutico) y se cuantificó colorimétricamente la actividad de fosfatasa alcalina, que se expresa bajo el control del promotor del factor transcripcional de NF-kb. Se evalúa el % actividad de NF-kb en efecto preventivo y terapéutico respecto a cultivos control de células con LPS y sin extracto añadido, que se consideran 100% de NF-kb. Resultados: La capacidad antiinflamatoria preventiva del extracto a 50 mg/l es del 25,5% (IC 95% 16,8-34,2) y el efecto terapéutico del 34,9% (IC 95% 25,3-44,4) para la misma concentración, no presentando actividad significativa a 10 mg/l. Conclusión: Se muestra una actividad de los polifenoles extraídos de los huesos de aceitunas, tanto preventivo de la inflamación como terapéutico de eliminación de la inflamación a través de la inhibición del factor NF-kB previamente activado por LPS a concentraciones de 50 mg/l de polifenoles que previamente se han mostrado seguras.

The aim of this study was to assess the anti-inflammatory activity of a polyphenolic extract from olive pits. Material and methods: The THP1-XBlue-CD14 (invivogen) cellular line, 80,000 cells/well, was incubated and inflammation (activation of NF-kb) was produced with 0.1 mg/mL of LPS (lipopolysaccharide from E. coli) for 24 hours. We assessed the presence of the extract (10 and 50 mg/L, biologically safe concentrations) for 2 hours at 37º C, before (preventive effect) and after (therapeutic effect) the proinflammatory activation, and the activity of alkaline phosphatase, which is expressed under the control of the NF-kb transcriptional factor, was quantified by colorimetry. The percentage of activity of NF-kb as preventive effect and therapeutic effect was assessed by comparing it to control cultures of cells with LPS and without extract, which are considered 100% of NF-kb. Results: The preventive anti-inflammatory capacity of the extract at 50 mg/L was 25.5% (95% CI: 16.8-34.2) and the therapeutic effect 34.9% (95% CI: 25.3-44.4) for the same concentration, without any significant activity at 10 mg/L. Conclusion: An activity of polyphenols extracted from olive pits is shown, both in preventing inflammation and therapeutically eliminating inflammation through inhibition of NF-kB factor previously activated by LPS at concentrations of 50 mg/L of polyphenols, which previously haven been shown to be safe.