Background The aim of this study was to compare pre-mortem histopathology findings in rats with and without traumatic brain injury (TBI) and its prospective application in forensic medicine. Methods This study involved 12 rats with 6 rats for each treatment group. This type of study is a laboratory experimental study with two independent groups design. The first group were rats that did not experience TBI. The second group was a group of rats with TBI. The subjects of this study were Rattus norvegicus rats, adult males, 4-8 weeks old, weighing 150-200 grams. On the 8th day after the rats experienced traumatic brain injury, the rats were then euthanized using the cervical dislocation method, after euthanasia the rats were given craniotomy and brain tissue was taken for histopathology examination. Results The description of histopathology changes in the brain organs in the group of rat without TBI found that neuron cells looked normal although there were also degeneration (21.16 ± 2.56/FV), necrosis (5.75 ± 0.98/FV), apoptosis (2.91 ± 0.80/FV), congestion ( 0.91 ± 0.49/FV), inflammatory cells (4.58 ± 1.15/FV) and hemorrhage (2.41 ± 1.11/FV). Changes in the rat traumatic brain injury group showed a lot of damage to neuron cells in the form of degeneration (48.41 ± 3.27/FV), necrosis (36.66 ± 2.89/FV), apoptosis (18.91 ± 1.24/FV), congestion (2.50 ±0.31/FV), inflammatory cells (11.41 ± 1.71/FV) and hemorrhage (10.08 ± 2.17/FV). Based on the results of statistical analysis, it can be seen that in all parameters there is a significant difference (p ≤ 0.001). Conclusions The premortem histopathology findings in rats with and without TBI which can be used for the benefit of forensic medicine in determining whether TBI is present or not. It is necessary to look more closely at the histopathology changes in the form of necrosis, apoptosis and hemorrhage.