We evaluated the protein levels of neuron-specific enolase (NSE) and S-100beta in serum and cerebrospinal fluid (CSF) in an animal model of acute spinal cord injury and ascertained their relevance. Spinal cord injury was induced at the T8 level in rats. Enzyme-linked immunosorbent assay was used to measure the protein levels of NSE and S-100beta in both serum and CSF at different time points (30 min, 2 h, 6 h, 12 h and 24 h after induction of spinal cord injury). There existed a significant correlation between neurological deficits and the severity of spinal cord injury (p<0.05). Compared with the control group, the protein levels of NSE and S-100beta in serum and CSF significantly increased from 2 h after injury (p<0.05) and reached a maximum at 6 h. Within a certain time window, the protein levels of NSE and S-100beta in serum and CSF were closely related to the severity of injury level (p<0.05). The protein levels of NSE and S-100beta in serum and CSF significantly increased after experimental spinal cord injury in a time-dependent manner and thus may be considered specific biomarkers for acute spinal cord injury.