Neonatal hypoxic‐ischaemic encephalopathy (HIE) refers to brain damage caused by intra‐uterine distress and asphyxia/hypoxia during the perinatal and neonatal periods. MicroRNA (MiR)‐214‐3p plays a critical role in cell growth and apoptosis. The aim of this study was to investigate the expression and role of miR‐214‐3p in neonatal HIE development, and to explore the underlying mechanisms. The expression of miR‐214‐3p was significantly down‐regulated, while that of Slc8a1, a direct target of miR‐214‐3p, was significantly up‐regulated, in the brain tissue of neonatal HIE rats. The over‐expression of miR‐214‐3p promoted the proliferation and inhibited the apoptosis of neurones, while its down‐regulation had the opposite effect. Our results indicate that miR‐214‐3p expression was down‐regulated in neonatal HIE rats, and the up‐regulation of miR‐214‐3p expression protected against HIE development by inhibiting neuronal apoptosis.