Protection of glutamine: The NF-κB/MLCK/MLC2 signaling pathway mediated by tight junction affects oxidative stress, inflammation and apoptosis in snakehead (Channa argus).

Lipopolysaccharide (LPS) destroys intestinal mechanical barrier and causes apoptosis by triggering oxidative stress and inflammatory responses. Glutamine (Gln) can maintain normal intestinal function under various stressed or pathological conditions. Thereby, this study aims to evaluate the protection of glutamine on intestinal health of snakehead (Channa argus), specifically regarding the NF-κB/MLCK/MLC2 signaling pathway mediated by tight junction affecting oxidative stress, inflammation and apoptosis. In this work, a model of intestinal tight junction injury in intestine of snakehead was constructed by injecting 4 mg/mL LPS into anus for 96 h. Before constructing the model, fish were treated with different levels of alanyl-glutamine (Ala-Gln) (0 %, 0.3 %, 0.6 %, 0.9 %, 1.2 % and 1.5 %) for 56 days. Microstructure and ultra microstructure showed that LPS-induced obvious intestinal damage and tight connection destruction, while Gln effectively alleviated these phenomena. In addition, results also showed that Gln can effectively inhibit LPS-induced damage to intestinal tight junction (zo-1, occludin, claudin5, claudin1, nf-κb p65, mlck and mlc2), alleviate oxidative stress (nrf2, sod, gsh, gpx and cat), ameliorate intestinal inflammation (tnf-α, il-1β, il-8, tlr5 and tlr2), thereby reduce apoptosis (p38mapk, caspase9, caspase8, caspase3 and bax). Crucially, the above results were related to NF-κB/MLCK/MLC2 signaling pathway mediated by tight junction. In conclusion, Gln has a good protective effect on LPS-induced intestinal injury in northern snakehead, providing a new perspective for regulating fish intestinal health.