OBJECTIVES Necrotizing enterocolitis (NEC) frequently results in significant morbidity and mortality in premature infants. Others reported that mice deficient in pulmonary surfactant protein-A (SP-A) born and raised in a nonhygienic environment succumb to significant gastrointestinal tract pathology, and enteral administration of purified SP-A significantly reduced mortality. We hypothesized that oral administration of purified SP-A can ameliorate pathology in an experimental model of neonatal NEC. METHODS Experimental NEC was induced in newborn Sprague-Dawley rat pups by daily formula gavage and intermittent exposure to hypoxia. Purified human SP-A (5 μg/day) was administered by oral gavage. After 4 days, surviving pups were sacrificed, and intestinal pathology was assessed by histological examination of distal terminal ileal sections. Intestinal levels of inflammatory cytokines (IL-1β, IFN-γ and TNF-α) were assessed by ELISA and levels of toll-like receptor 4 (TLR4) by western analysis. RESULTS 61% of the gavaged rat pups that survived to day 4 met the criteria for experimental NEC after hypoxia while treatment with SP-A significantly reduced mortality and assessment of NEC. Intestinal levels of pro-inflammatory cytokines were significantly increased in pups exposed to hypoxia. Administration of SP-A to pups exposed to hypoxia significantly reduced IL-1β and TNF-α levels, but had little effect on elevated levels of IFN-γ. SP-A treatment of hypoxia-exposed pups significantly reduced expression of intestinal TLR4, key in NEC pathogenesis. CONCLUSIONS In a rat model of experimental neonatal NEC, oral administration of SP-A reduces intestinal levels of pro-inflammatory cytokines and TLR4 protein, and ameliorates adverse outcomes associated with gastrointestinal pathologies.