Antioxidant and anti-inflammatory activities ofPinus radiatabark extract in salmonid cell lines

Abstract

A fish meal supply shortage is limiting aquaculture development. Currently, plant-based proteins, such as soya bean meal, are being used as an alternative protein source, despite that such a diet can adversely affect fish, such as by inducing an inflammatory response. A possible solution is to include dietary additives in farm diets to counteract negative effects. One such solution originates from pine bark extracts, which present bioactive properties. In this study, the antioxidant and anti-inflammatory properties of Pinus radiata bark extracts were evaluated for the first time in a salmonid cell line. This extract chemically demonstrated antioxidant activity through 2,2-diphenyl-1-picrylhydrazyl (DPPH = 58.4 ± 1.1%) and ferric ion reducing antioxidant power (FRAP = 575 ± 17 mgEqFe(II)·g extract−1) assays. Additionally, the extract showed high flavonoid and phenolic compound contents. Up to 100 mg mL−1, the P. radiata extract showed no cytotoxicity in the CHSE-214 salmonid embryo cell line. Moreover, the antioxidant activity of the extract (50 μg mL−1) was evaluated by a dichlorofluorescein (DCFH) assay in the SHK-1 salmon cell line challenged with an oxidant stimulus (H2O2), showing 58.9% activity. The extract also protected DNA from oxidative damage, as observed through a comet assay. When assessing anti-inflammatory properties in an in vitro inflammation model, the extract significantly reduced the relative expression of the pro-inflammatory cytokines interleukin-1β (IL-1β), tumour necrosis factor-α (TNF-α) and interleukin-8 (IL-8) and of the inducible cyclooxygenase-2 (COX-2) enzyme. These results suggest a potential application of P. radiata bark extract in functional foods in aquaculture.