Abstract: Objective To explore the levels and clinical significance of serum thrombospondin 1(TSP1), thrombospondin 2(TSP2), neuron-specific enolase(NSE)and S-100 β before and after trepanation and drainage in chronic subdural hematoma(CSDH). Methods A total of 82 patients with CSDH admitted to our hospital given trepanation and drainage were selected as CSDH group(further subdivided into mild group and moderate group according to the degree of brain injury), and another 55 healthy examiners in the same time period were selected as control group. Serum levels of TSP1, TSP2, NSE and S-100β were determined by enzyme-linked immunosorbent assay. The relationships between trepanation and drainage, cerebral injury, postoperative recurrence and their expression levels were analyzed. Results The levels of serum TSP1, TSP2, NSE and S-100β in CSDH patients were significantly higher than those of controls(P<0.05). The levels of serum TSP1, TSP2, NSE and S-100β in mild group were significantly lower than those in moderate group(P<0.05). The levels of serum TSP1, TSP2, NSE and S-100β in CSDH patients at 7 d after surgery - were significantly lower than those before surgery(P<0.05). The levels of serum TSP1, TSP2, NSE and S-100β in patients with postoperative recurrence were significantly higher than those without(P<0.05). The changes of serum TSP1 before and after trepanation and drainage in CSDH patients were positively correlated with the changes of serum NSE and S-100β(r=0.423, 0.389, P<0.05), and the changes of serum TSP2 before and after trepanation and drainage in CSDH patients were positively correlated with changes of serum NSE and S-100β(r=0.418, 0.392, P<0.05). Conclusion The levels of serum TSP1, TSP2, NSE and S-100β in CSDH patients are decreased after trepanation and drainage, and are closely related to the brain injury degree and outcomes.