Abstract: Objective To investigate the effect of erythropoietin (EPO) on the expression of MMP-2 in hippocampus of neonatal rats after hypoxic-ischemic brain damage (HIBD), and the mechanism of its neuroprotective effect. Methods Neonatal Sprague-Dawley rats of 7 days old were randomly divided into three groups (n = 48 in each group): sham-operated group, HIBD group and EPO treated group, then each group was further divided into four subgroups (n = 12) based on different time points following the injection of medication ( 6 h, 24 h, 3 d, 7 d). The expression of MMP-2 in hippocampus was determined by immunohistochemistry and real-time fluorescent quantitative PCR method. Results Immunohistochemistry: MMP-2 has a small amount of expression in the hippocampus of the sham-operated group, and at each time point, there is no statistically significant difference (P > 0.05); The expression of MMP-2 in HIBD group and EPO group all show a trend of increase, and peaked at 7 d, the differences between each time point in two groups are statistically significant (P < 0.05); Compared with control group, the difference in each time point of the other two groups showed significance (P < 0.05) in addition to the 6 h point, and there is significant difference at the 7 d point between EPO group and HIBD group (P < 0.05). RT-qPCR: The gene expression of MMP-2 in control group presents a trend of increase, but there is no significant difference at different time points (P > 0.05). Gene expression in HIBD group at 24 h and 7 d points showed twin peaks, and the peak is higher at the 7 d point, but without difference (P > 0.05). MMP-2 expression of EPO group presents a trend of increase, and differences are significant between each time point (P < 0.05); At each time point, the expression of MMP-2 mRNA in both HIBD group and EPO group is extremely high than that in sham-operated group (P<0.05). Compared with the HIBD group, the expression of MMP-2 mRNA at 24 h of EPO group decreased, but it is significantly higher at the time of 7 d (P < 0.05). Conclusion Erythropoietin may upregulate the expression of MMP-2 in the delayed phase of HIBD, which may be one of the mechanisms for protecting HIBD.