Heme oxygenase-1 (HO-1) is an inducible rate-limiting enzyme for heme degradation. Here, we studied the HO-1 expression in an open-skull weight-drop-induced traumatic brain injury, with a focus on the early phase, most amenable to therapy. In normal rat brains of our study, HO-1+ cells were rarely observed. Significant parenchymal accumulation of HO-1+ non-neuron cells was observed 18 h post-traumatic brain injury and increased continuously during the investigating time. We also observed that the accumulated HO-1+ non-neuron cells were mainly distributed in the perilesional areas and showed activated microglia/macrophage phenotypes with ramified or amoeboid morphologic characteristics. Further double-labeling experiments showed that most HO-1+ non-neuron cells coexpressed CD68 and CD163, but not glial fibrillary acid protein. Our data suggest that HO-1 expression defines a subtype of activated microglia/macrophages involved in the early processes following traumatic brain injury.