Protective Effect of SIRT1-mediated NF-κB Signal Pathway against Necrotizing Enterocolitis in Neonatal Mice

Abstract

Objective To discover the mechanism of Sirtuin1 (SIRT1)-mediated NF-κB pathway in the protection of necrotizing enterocolitis (NEC) in neonatal mice. Methods Neonatal mice were treated with EX527 (an inhibitor of SIRT1) and/or pyrrolidine dithiocarbamate (PDTC, an inhibitor of NF-κB). The survival rate of mice was recorded. Hematoxylin & eosin (HE) staining was performed to observe the pathological changes of intestines. Furthermore, western blotting, Enzyme-linked immuno sorbent assay (ELISA), and real-time quantitative polymerase chain reaction (qRT-PCR) were conducted to measure the protein and gene expression, while corresponding kits to detect the levels of oxidative stress indicators. Results PDTC increased the survival rate of NEC mice. When compared with the NEC + EX527 + PDTC group, the histological score of NEC was higher in the NEC + EX527 group but was lower in the NEC + PDTC group. SIRT1 expression in the intestines of NEC mice was down-regulated, with an increase in p65 nuclear translocation. Also, MDA increased and GPx decreased in the intestines of NEC mice, with the up-regulations of IL-6, IL-1β and TNF-α, as well as the down-regulations of ZO-1, occludin and claudin-4 in the intestines. However, changes above could be improved by PDTC, which would be further reversed by EX527. Conclusion SIRT1 could mitigate inflammation and oxidative stress response and improve the intestinal permeability by mediating NF-κB pathway, playing an important role in the alleviation of NEC.