In this study, we investigated the time-dependent expression of IL-1β, COX-2, MCP-1 mRNA after incised wounds in rabbit skin using real-time fluorescent quantitative PCR. The tested wound ages were distributed as following: <0.5 h, 0.5 h, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 8 h, 12 h, 24 h, 2 d, 3 d, and 7 d. The expressions of three markers in postmortem wounds were determined. Comparison of each wound age with control group, expression of IL-1β mRNA showed that the significant increase occurred at <0.5 h ( p < 0.01), and the peak level at 2 h. The expression was almost normalized at 2 d. But for COX-2 and MCP-1 mRNA, the significant increase occurred at 1 h for COX-2 mRNA, and 3 h for MCP-1 mRNA. The expression peak levels were at 3 h and 5 h, and were almost normalized at 3 d and 7 d, respectively. There was no significant increase in all postmortem samples for IL-1β, COX-2, MCP-1 mRNA compared with control group. Thus, the results of these cytokines and enzyme significant increase at different early wound ages implied that the combined investigation could make wound age determination more objective and accurate. Moreover, the three markers could also be used to distinguish the supravital injuries.