Lactoferrin Alleviates Lipopolysaccharide-Induced Infantile Intestinal Immune Barrier Damage by Regulating an ELAVL1-Related Signaling Pathway

Abstract

As the most important intestinal mucosal barrier of the main body, the innate immune barrier in intestinal tract plays especially pivotal roles in the overall health conditions of infants and young children; therefore, how to strengthen the innate immune barrier is pivotal. A variety of bioactivities of lactoferrin (LF) has been widely proved, including alleviating enteritis and inhibiting colon cancer; however, the effects of LF on intestinal immune barrier in infants and young children are still unclear, and the specific mechanism on how LF inhibits infantile enteritis by regulating immune signaling pathways is unrevealed. In the present study, we firstly performed pharmacokinetic analyses of LF in mice intestinal tissues, stomach tissues and blood, through different administration methods, to confirm the metabolic method of LF in mammals. Then we constructed in Vitro and in Vivo infantile intestinal immune barrier damage models utilizing lipopolysaccharide (LPS), and evaluated the effects of LF in alleviating LPS-induced intestinal immune barrier damage. Next, the related immune molecular mechanism on how LF exerted protective effects was investigated, through RNA-seq analyses of the mouse primary intestinal epithelial cells, and the specific genes were analyzed and screened out. Finally, the genes and their related immune pathway were validated in mRNA and protein levels; the portions of special immune cells (CD4+ T cells and CD8+ T cells) were also detected to further support our experimental results. Pharmacokinetic analyses demonstrated that the integrity of LF could reach mice stomach and intestine after oral gavage within 12 h, and the proper administration of LF should be the oral route. LF was proven to down–regulate the expression levels of inflammatory cytokines in both the primary intestinal epithelial cells and mice blood, especially LF without iron (Apo-LF), indicating LF alleviated infantile intestinal immune barrier damage induced by LPS. And through RNA-seq analyses of the mouse primary intestinal epithelial cells treated with LPS and LF, embryonic lethal abnormal vision Drosophila 1 (ELAVL1) was selected as one of the key genes, then the ELAVL1/PI3K/NF-κB pathway regulated by LF was verified to participate in the protection of infantile intestinal immune barrier damage in our study. Additionally, the ratio of blood CD4+/CD8+ T cells was significantly higher in the LF-treated mice than in the control mice, indicating that LF distinctly reinforced the overall immunity of infantile mice, further validating the strengthening bioactivity of LF on infantile intestinal immune barrier. In summary, LF was proven to alleviate LPS-induced intestinal immune barrier damage in young mice through regulating ELAVL1-related immune signaling pathways, which would expand current knowledge of the functions of bioactive proteins in foods within different research layers, as well as benefit preclinical and clinical researches in a long run.

Targeting AHR Increases Pancreatic Cancer Cell Sensitivity to Gemcitabine through the ELAVL1-DCK Pathway

“…ELAVL1 is known to be involved in various cell signalling pathways, such as MAPK and JNK [48], as well as ferroptosis activation [52]. It is also involved in various systems, such as innate immune barriers in infants [53] and the overall immune system [45,53]. Similarly, AHR is also involved in various cell signalling pathways, such as epidermal growth factor family pathways [54] and xenobiotic metabolism [55].…”

Milk: A Natural Guardian for the Gut Barrier

“…Moreover, it decreased intestinal permeability, thereby bolstering the integrity of the intestinal barrier and offering protection against bacterial infections . Furthermore, LF can reduce the expression of inflammatory markers such as IL-1β, IL-6, TNF-α, and IFN-γ while concurrently increasing the ratio of CD4/CD8 T cells in the bloodstream of LPS-infected mice . From the above, LF has shown great potential for gut barrier protection from different aspects.…”

“…69 Furthermore, LF can reduce the expression of inflammatory markers such as IL-1β, IL-6, TNFα, and IFN-γ while concurrently increasing the ratio of CD4/ CD8 T cells in the bloodstream of LPS-infected mice. 67 From the above, LF has shown great potential for gut barrier protection from different aspects.…”

Lactoferrin, the Moonlighting Protein of Innate Immunity

“…As an ancestral guardian, the primary defense activities of Lf against pathogens and their sequelae have been dissected in different studies in the present Special Issue. Li and colleagues have investigated the beneficial effect of Lf administration in a model of lipopolysaccharide (LPS)-induced intestinal immune barrier damage [17]. The innate immune barrier of the intestinal tract plays a crucial role in human health, especially in infants and young children who have an immature immune system [18].…”

“…In addition, the ratio of blood CD4+/CD8+ T cells was significantly higher in the Lf-treated mice than in the control mice. Lactoferrin was therefore proven to alleviate LPS-induced intestinal immune barrier damage in young mice through the regulation of ELAVL1-related immune signaling pathways, which would expand the current knowledge of the functions of bioactive proteins in food within different levels of research, as well as preclinical and clinical research in the long run [17].…”