Objective To investigate the clinical value of the brain injury markers including neuron specific enolase (NSE) and brain active peptide 100-β (S-100 β) in the acute toadstools poisoning patients and to provide evidence-based support for clinical treatment and prognosis. Methods 134 poisoning patients were divided into four groups according to PSS scores. The serum NSE and S-100β levels were measured by chemiluminescence immunoassay. Results The serum NSE and S-100β levels in poisoning groups were higher than that of normal control groups (P<0.05). The serum NSE and S-100β levels were correlated with the degree of poisoning. Conclusion The serum NSE and S-100β levels could reflect the brain injury degree of toadstools poisoning patients. It has certain significance in guiding the treatment and prognosis.