Abstract: Objective To study the clinical effect of neonatal necrotizing enterocolitis (NEC) with erythropoietin and the influences of inflammation and toll like receptor(TLR)4/NF-κB signal pathway. Methods A total of 94 consecutives as NEC from August 2016 to August 2018 were enrolled and divided into control group (n=42) and observation group (n=52) ; those in control group received standard medical treatment plan and those in observation group adopted recombinant EPO intramuscular injection (dosage of 200 IU/kg, twice per week) at the same time for the course of one week. Then to compare differences of complications and mortality incidences, serum levels of TNF-α and IL-6,TLR-4 and NF-κB mRNA before and after treatment. Results In the control group, 21 patients had complications, including 7 cases sepsis, 6 cases acute intestinal perforation, 6 cases infectious peritonitis, and 2 cases other cases. There were 15 cases in the observation group, including 5 cases sepsis and 5 cases acute intestinal perforation, 4 cases infectious peritonitis, 1 other case, The complications and mortality incidences in observation group were both significantly lower than control group (all P＜0.05). What's more, the levels of TNF-α and IL-6 after treatment in observation group were both lower than before, and less than control group (all P＜0.05). The levels of TLR-4 and NF-κB mRNA after treatment in observation group were both lower than before, and less than control group, too (all P＜0.05). Conclusion It can reduce inflammation TNF-α and IL-6 levels by TLR-4/NF-κB signal pathway with EPO in order to improve NEC clinical symptoms and prognosis.