Objective To investigate the dynamic expression and clinical significance of myoglobin, creatine kinase and inflammatory mediators in the serum of patients with multiple trauma. Methods From May 2013 to March 2015, 56 patients with multiple trauma admitted in EICU were recruited in the study. According to the injury severity, 56 patients were divided into a mild trauma group, a medium trauma group and a severe trauma group. The subjects were further divided into a MODS group and a non-MODS group based on multiple organ dysfunction syndrome (MODS) criteria. Twenty healthy adults undergoing physical examination were recruited as control. Serum myoglobin, creatine kinase, IL-6 and TNF-α levels were measured in the multiple trauma patients (1st day, 3rd day, 7th day and 14th day) and the controls. Results Compared with the controls, the serum levels of myoglobin, creatine kinase, IL-6 and TNF-α in the patients with multiple trauma increased significantly from 1st to 14th day after injury (allP＜0.05). Serum myoglobin, creatine kinase, IL-6 and TNF-α levels on 3rd day after injury reached the peak, then decreased gradually in the mild, medium, and severe trauma groups, among which the changes of serum myoglobin, creatine kinase, IL-6 and TNF-α levels were significant on 3rd day compared with other timepoints (allP＜0.05). On 1st day after injury, serum levels ofmyoglobin, creatine kinase, IL-6 and TNF-α also differed significantly between the MODS group and non-MODS group (allP＜0.05). The AUCs of myoglobin, IL-6 and TNF-α for predicting MODS were 0.527-0.817, 0.641-0.890, and 0.197-0.544, respectively. Conclusions The dynamic changes of serum myoglobin, creatine kinase, IL-6 and TNF-α in patients with multiple trauma are correlated well with the injury severity and prognosis. Serum myoglobin, IL-6 and TNF-α levels may be good markers to predict secondary MODS in multiple trauma patients.