Modern approaches to the treatment of traumatic brain injury (TBI) focus on alleviating life-threatening conditions. However, background glial activation processes occurring throughout the brain are often overlooked and may contribute to long-term damage and cognitive decline in patients with TBI. Currently, there is increasing evidence to support the influence of endogenous lipid messengers such as N-acylethanolamines fatty acids in neuropathologies. Synaptamide (N-docosahexaenoylethanolamine) plays an important role in brain repair after neurotrauma due to its significant anti-inflammatory, gliotropic and neuroprotective effects. The aim of this study was to investigate the processes of reactive astrogliosis occurring in two brain regions (cortex and thalamus) following cerebral cortex injury and synaptamide therapy (10 mg/kg/day, 7 days) in mice. Synaptamide was shown to reduce the expression of brain damage biomarkers in the blood, inhibit the development of reactive astrogliosis, and trigger the activation of anti-inflammatory A2 type reactive astrocytes, with the synthesis of the main brain neurotrophins (Clcf1, BDNF) in vitro and in vivo. The therapeutic efficacy of synaptamide demonstrated in this study makes it a promising drug for the treatment of TBI.