Abstract
Chinese olive (Canarium album L.), a rich source of polyphenols, can be used as a functional food ingredient. We previously showed that the ethyl acetate fraction of this extract (CO-EtOAc) is an effective anti-inflammatory agent. Therefore, here, we aimed to screen the bioactive fractions extracted from CO-EtOAc using different isolation techniques, and purify the bioactive compounds based on their cytotoxic and anti-inflammatory abilities. CO-EtOAc was fractionated using silica gel and Sephadex column chromatography, and the active compounds were isolated and purified by high-performance liquid chromatography (HPLC). The structures of the resulting compounds were identified using proton nuclear magnetic resonance (NMR) spectra. Activity-directed fractionation and purification were used to identify the following active compounds with anti-inflammatory effects using lipopolysaccharide (LPS)-stimulated mouse macrophages: sitoindoside I, amentoflavone, tetrahydroamentoflavone and protocatechuic acid. For the first time, sitoindoside I and tetrahydroamentoflavone were isolated from Chinese olive, and the anti-inflammatory compounds of CO-EtOAc were identified, suggesting its potential for used as a health food ingredient.
Highlights
A plethora of accumulating evidence has proven that inflammation plays a central role in the pathogenesis of various disorders such as cancer, diabetes, obesity, and cardiovascular complications.Controlling inflammation is, of major importance in treating chronic inflammation-associated illnesses [1]
The RAW264.7 macrophage cell line is extensively used as a cell model in the study of the signal transduction pathway upon activation of pro-inflammatory mediators [3], including nuclear factor-kappaB (NF-κB), activator protein-1 (AP-1), interleukin-1 beta (IL-1β), IL-6, IL-8 and nitric oxide (NO) [4,5,6]
The results showed that sitoindoside I, amentoflavone, and protocatechuic acid at concentrations of concentrations of μg/mL
Summary
A plethora of accumulating evidence has proven that inflammation plays a central role in the pathogenesis of various disorders such as cancer, diabetes, obesity, and cardiovascular complications.Controlling inflammation is, of major importance in treating chronic inflammation-associated illnesses [1]. The RAW264.7 macrophage cell line is extensively used as a cell model in the study of the signal transduction pathway upon activation of pro-inflammatory mediators [3], including nuclear factor-kappaB (NF-κB), activator protein-1 (AP-1), interleukin-1 beta (IL-1β), IL-6, IL-8 and nitric oxide (NO) [4,5,6]. Of these mediators, NO serves as an important player in. Inhibiting NO production in LPS-stimulated RAW264.7 cells can be adopted to screen various anti-inflammatory drugs. More attention has turned to the development of new drugs with a potency that will inhibit NO production; this production can be detected using a fast and simple colorimetric assay, the Griess reaction-based method [12,13]
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