Oxidized fish oil injury stress in Megalobrama amblycephala: Evaluated by growth, intestinal physiology, and transcriptome-based PI3K-Akt/NF-κB/TCR inflammatory signaling

Lipids are essential nutrients for animal. Oxidized lipid might induce injury stress for fish. Here we conducted a 12-week rearing experiment with diets containing 0, 2, 4, and 6% oxidized fish oil (6F, 4F2OF, 2F4OF, and 6OF) to describe the oxidative impairment mechanism on teleost fish blunt snout bream, Megalobrama amblycephala. Results were evaluated by growth performance, intestinal physiology, and transcriptome-based PI3K-Akt/NF-κB/TCR inflammatory signaling. From the results, 6OF reduced growth performance with increased FCR and reduced FBW, WGR and SGR compare with 6 F. Meanwhile, oxidized fish oil treatments also increased antioxidant enzyme activity, suggesting an impaired physiological condition. The plasmatic antioxidant enzyme activity of T-SOD, GSH-Px, ASAFR, concentration of MDA and cortisol were significantly increased in 6OF, while GSH concentration was decreased. Histological ultrastructure revealed the integrity of mid-intestinal cells and villus were destroyed in 6OF. Moreover, transcriptomic analysis revealed PI3K-Akt/NF-κB/TCR inflammatory signaling were active to oxidized fish oil stress. We verified the expression of twelve key genes related to this signaling by RT-PCR, which revealed TLR2, PI3K, Akt, NF-κB, MHCII-β, TCR-α, TGF-β, TNF-α, IL-6, IL-1β, GPx1 and GSTm were all activated under 6OF stimulation. We found that oxidized fish oil may induce oxidative stress, destroy intestinal integrity, produce free radical, dysregulate lipid metabolism and oxidative balance, reversely affect the physiological adaptation, and eventually lead to growth inhibition. This study revealed the mechanism of PI3K-Akt/NF-κB/TCR inflammatory signaling in M. amblycephala under oxidized fish oil stress, which may help to understand the complex regulation involved in lipid oxidative stress resistance.