Journal
FISH & SHELLFISH IMMUNOLOGY
Volume 35, Issue 3, Pages 933-940Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.fsi.2013.07.004
Keywords
Eicospentaenoic (EPA) or docosahexaenoic (DHA); Larmichthys crocea; In vitro; Macrophage; Immune response
Funding
- National Nature Science Fund of China - Research Funding Program I: Effects of fatty acids on the physiology and immune responses of large yellow croaker [30871930, C:120305]
- Research Funding Program II: Study on the phospholipid metabolism of large yellow croaker larvae [31172425, C:1905]
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Comparative effects of different concentrations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on immune responses of head-kidney macrophages isolated from large yellow croaker were studied in vitro. After exposing to serum-free medium for 1 day, cultured cells were incubated in medium supplemented with graded levels of EPA or DHA (0, 5, 25, 100, 200 and 1000 mu M, respectively) in the form of fatty acid bovine serum albumin (FA-BSA) complex for 12 h, 24 h and 36 h, respectively. Control samples were incubated in the absence of EPA or DHA (2% bovine serum albumin, BSA). Following stimulation, cell viability, lipid peroxidation, secretary phopholipase A(2) (sPLA(2)) and prostaglandin E-2 (PGE(2)) production as well as some immune parameters including phagocytosis, respiratory burst activity and interleukin 1 beta (IL-1 beta) production were determined. Results showed that EPA and DHA affected cell viability in dose-dependent and time-dependent manners. In particular, cell viability was significantly decreased after 24 h and 36 h incubation with 1000 mu M EPA or DHA (P < 0.05). Higher levels of EPA (200 and 1000 mu M) caused a significant increase in the production of malondialdehyde (MDA) (P < 0.05), while DHA did not significantly affect the MDA production. EPA significantly increased the intracellular superoxide anion synthesis which, on the contrary, was significantly reduced by DHA. Phagocytosis percentage (PP) values were significantly higher in treatments with 5 mu M DHA (P < 0.05), but significantly decreased by 200 and 1000 mu M EPA and DHA compared to the control group (P < 0.05). Decreased PGE2 production was produced by cells treated with relatively low doses of EPA or DHA. When high levels of stimulants (1000 mu M EPA or DHA) were used, PGE(2) levels were elevated and reached a significant level (P < 0.05). Both EPA and DHA significantly inhibited the production of sPLA(2), where DHA exerted the more potent inhibitory effects than EPA. No pronounced effect was observed on IL-1 beta production among all the treatments, and IL-1 beta level in cell culture supernatant was fairly low (only approximately 6 pg/ml). Those findings suggested that EPA and DHA could influence the immunity and physiological conditions of macrophages from head kidney of large yellow croaker in vitro. (C) 2013 Published by Elsevier Ltd.
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