The Protective Effects of Iron Free Lactoferrin on Lipopolysaccharide-Induced Intestinal Inflammatory Injury via Modulating the NF-κB/PPAR Signaling Pathway

Research evidence shows that effective nutritional intervention could prevent or reduce intestinal inflammatory injury in newborn infants. Iron free lactoferrin (apo-LF), one of the main types of lactoferrin (LF), is a bioactive protein in milk that plays a vital role in maintaining intestinal health. The potential mechanism by which apo-LF modulates intestinal inflammation is, however, still unclear. In the study we first explored key genes and pathways in vitro by transcriptome date analysis and then validated them in vivo to reveal the underlying molecular mechanism. The results showed that apo-LF pretreatment effectively inhibited lipopolysaccharide (LPS)-induced primary intestinal epithelial cells (IECs) inflammation in the co-culture system (primary IECs and immune cells), which was specifically manifested as the reduction of the concentration of TNF-alpha, IL-6 and IL-1 beta and increased the concentration of IFN-gamma. In addition, transcriptome data analysis revealed that the key pathway for apo-LF to exert anti-inflammatory effects was the NF-kappa B/PPAR signaling pathway. Further validation was performed using western blotting in colonic tissues of young mice and it was found that the major proteins of NF-kappa B signaling pathway (NF-kappa B, TNF-alpha and IL-1 beta) were inhibited by apo-LF and the target proteins of PPAR signaling pathway (PPAR-gamma and PFKFB3) were activated by apo-LF. Taken together, this suggests that apo-LF has a protective effect against LPS-induced intestinal inflammatory injury via modulating the NF-kappa B/PPAR signaling pathway, which provides new insights for further anti-inflammatory study of apo-LF.