Abstract: Objective　To explore the relationship of serum ubiquitin carboxy terminal hydrolase L1 (UCH-L1) and glial fibrillary acidic protein (GFAP) with brain injury in preterm infants. Methods　A total of 130 premature infants with gestational age <34 weeks from August 2014 to October 2016 were recruited. Blood samples were collected at 6 h and 72 h after birth. The levels of serum UCH-L1 and GFAP were detected by ELISA method. According to the results of cranial ultrasound and MRI examination, the premature infants were divided into white matter damage (WMD) group, periventricular intraventricular hemorrhage (PVH-IVH) group, and no brain injury group. The levels of serum UCH-L1 and GFAP in preterm infants between the three groups, mild to severe brain injury were compared. Results　At 6 h and 72 h after birth, the levels of serum UCH-L1 and GFAP among no brain injury group, PVH-IVH group and WMD group were significantly different (all P <0.001). The level of serum UCH-L1 and GFAP were the highest in the WMD group and the lowest in no brain injury group at both 6 h and 72 h after birth. The levels of serum UCH-L1 at 72 h after birth were significantly lower than those at 6 h after birth in PVH-IVH group and WMD group, while the levels of serum GFAP at 72 h after birth were significantly higher than those at 6 h after birth in both of the two groups (all P<0.05). The levels of serum UCH-L1 and GFAP in severe PVH-IVH group and severe WMD group were significantly higher than those in the mild group at 6 h and 72 h after birth (all P<0.05). Conclusions　The levels of serum UCH-L1 and GFAP in preterm infants can be used as sensitive markers for early evaluation of brain injury, which can help determine the severity of brain injury in preterm infants.