Abstract
Context: Interest has recently renewed in using Lolium multiflorum Lam. (Poaceae) (called Italian ryegrass; IRG) silage as an antioxidant and anti-inflammatory diet.Objective: This study investigated the antioxidant, anti-inflammatory and anti-septic potential of IRG silage and identified the primary components in IRG active fractions.Materials and methods: Total 16 fractions were separated from the chloroform-soluble extract of IRG aerial part using Sephadex LH-20 column before HPLC analysis. Antioxidant and anti-inflammatory activities of the fractions at doses of 0–100 μg/mL were investigated using various cell-free and cell-mediated assay systems. To explore anti-septic effect of IRG fractions, female ICR and BALB/c mice orally received 40 mg/kg of phenolic acid and flavonoid-rich active fractions F7 and F8 every other day for 10 days, respectively, followed by LPS challenge.Results: The active fractions showed greater antioxidant and anti-inflammatory potential compared with other fractions. IC50 values of F7 and F8 to reduce LPS-stimulated NO and TNF-α production were around 15 and 30 μg/mL, respectively. Comparison of retention times with authentic compounds through HPLC analysis revealed the presence of caffeic acid, ferulic acid, myricetin and kaempferol in the fractions as primary components. These fractions inhibited LPS-stimulated MAPK and NF-κB activation. Supplementation with F7 or F8 improved the survival rates of mice to 70 and 60%, respectively, in LPS-injected mice and reduced near completely serum TNF-α and IL-6 levels.Discussion and conclusion: This study highlights antioxidant, anti-inflammatory and anti-septic activities of IRG active fractions, eventually suggesting their usefulness in preventing oxidative damage and inflammatory disorders.
Highlights
Diet is known to be an important factor in maintaining a healthy life
Monoclonal antibodies specific for cyclooxygenase 2 (COX-2), inducible nitric oxide synthase and b-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA)
Regarding the mechanisms by the IRG active fractions inhibit the LPS-mediated induction of nitric oxide (NO), tumournecrosis factor-a (TNF-a), inducible nitric oxide synthase (iNOS) and COX-2, the current findings suggest the involvement of NF-jB-mediated signalling
Summary
Diet is known to be an important factor in maintaining a healthy life. Increased interest has focused on the application of dietary components to prevent inflammatory damage and/or to control immune response induction (Kamiyama & Shibamoto 2012; Yamaura et al 2012; Sun et al 2016). Diets abundant in antioxidants and anti-inflammatory constituents are believed to be an effective means of improving the resistance to pathogens. The feeding of fresh forage (either as pasture plus a concentrate or as a silage-based total mixed ration) is believed to improve the quality and quantity of animal products rather than the concentrate-rich diets did. It was reported that the feeding of fresh forage increased animal health, even for monogastric animals (Cuevas Montilla et al 2011; Firuzi et al 2011). These beneficial effects of plant-based diets are considered to be related to the presence of bioactive metabolites such as phenolic acids and flavonoidic compounds
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