Protective effect of chitosan oligosaccharide against oxidative damage of peripheral blood mononuclear cells in dairy cows induced by diethylenetriamine/nitric oxide via NF-κB signalling pathway

Excessive production of nitric oxide (NO) during the lactation period of cows leads to oxidative stress and inflammatory reactions. Chitosan has enhanced effects on antioxidant and immune function, but the mechanism of this effect is unclear. The current study investigated the underlying mechanism of the protective effect chitosan oligosaccharides (COS) exert against oxidative damage of peripheral blood mononuclear cells (PBMCs) in dairy cows. PBMCs were allocated to receive one of treatments: CTR: control treatment without COS addition; COS40, COS80, COS160, and COS320 received 40, 80, 160, and 320 μg/mL of COS; PBMCs were then treated with 200 μmol/L diethylenetriamine/nitric oxide (DETA/NO) for 4 h. Compared to the CTR group, the cell viability of the COS pre-treated group increased with increasing COS concentration. The activities of antioxidant enzymes including superoxide dismutase, catalase, and glutathione peroxidase and the thioredoxin reductase content showed an upward trend in a dose-dependent manner and COS at a concentration of 160 μg/mL exhibited the strongest effect. COS decreased the content and gene expressions of interleukin-1β and tumour necrosis factor-α, the mRNA and protein expression and activity of the inducible nitric oxide synthase, and NO production compared with the values observed in the CTR group. The expression of genes and proteins related to the nuclear factor kappa-B (NF-κB) pathway also showed a downward trend. Based on these indicators, we concluded COS has a protective effect against oxidative damage caused by DETA/NO, and we speculated that COS may play a protective role by inhibiting the NF-κB pathway.Highlights In vitro experiments simulated changes in antioxidant and immune functions caused by increased NO in dairy cows’ PBMCs in vivo. We showed that chitosan has a protective effect against oxidative damage and a regulatory effect on immune function in vitro. The regulation of chitosan against oxidative stress and immune responses may be related to the NF-KB signalling pathway