Role of intestinal Hsp70 in barrier maintenance: contribution of milk to the induction of Hsp70.2.
BACKGROUND: Necrotizing enterocolitis (NEC) is a gastrointestinal disease of complex etiology resulting in devastating systemic inflammation and often death in premature newborns. We previously demonstrated that formula feeding inhibits ileal expression of heat shock protein-70 (Hsp70), a critical stress protein within the intestine. Barrier function for the premature intestine is critical. We sought to determine whether reduced Hsp70 protein expression increases neonatal intestinal permeability.
METHODS: Young adult mouse colon cells (YAMC) were utilized to evaluate barrier function as well as intestine from Hsp70-/- pups (KO). Sections of intestine were analyzed by Western blot, immunohistochemistry, and real time PCR. YAMC cells were sub-lethally heated or treated with expressed milk (EM) to induce Hsp70.
RESULTS: Immunostaining demonstrates co-localized Hsp70 and tight junction protein zona occludens-1 (ZO-1), suggesting physical interaction to protect tight junction function. The permeability of YAMC monolayers increases following oxidant injury and is partially blocked by Hsp70 induction either by prior heat stress or EM. RT-PCR analysis demonstrated that the Hsp70 isoforms, 70.1 and 70.3, predominate in WT pup; however, Hsp70.2 predominates in the KO pups. While Hsp70 is present in WT milk, it is not present in KO EM. Hsp70 associates with ZO-1 to maintain epithelial barrier function.
CONCLUSION: Both induction of Hsp70 and exposure to EM prevent stress-induced increased permeability. Hsp70.2 is present in both WT and KO neonatal intestine, suggesting a crucial role in epithelial integrity. Induction of the Hsp70.2 isoform appears to be mediated by mother's milk. These results suggest that mother's milk feeding modulates Hsp70.2 expression and could attenuate injury leading to NEC.
LEVEL OF EVIDENCE: Level III.
Pediatric surgery international
Animals; Animals, Newborn; Cytoprotection; HSP70 Heat-Shock Proteins; Intestinal Mucosa; Mice; Milk; Permeability; Protein Isoforms; Tight Junctions; Zonula Occludens-1 Protein
Breast milk; Heat shock protein (HSP); Inflammation; Intestinal barrier; Necrotizing enterocolitis (NEC); Tight junction
Rentea, R. M., Guo, Y., Zhu, X., Musch, M., Chang, E., Gourlay, D., Liedel, J. Role of intestinal Hsp70 in barrier maintenance: contribution of milk to the induction of Hsp70.2. Pediatric surgery international 34, 323-330 (2018).