Aim: Neurodegeneration caused by the axonal injury is a widely seen phenomenonin spinal cord and traumatic brain injuries. Due to the disintegration of the synapticconnection neurotrophic factors could not be transported retrogradely towards thecell body and the deprivation of the trophic factors lead to the degeneration and deathof the injured neuron. Rifampicin is an antibiotic exhibiting several neuroprotectivefunctions in various neurodegenerative conditions. Here we aim to investigate theacute neuroprotective effect of rifampicin in primary cortical neuron culture in whichneurons are axotomized by laser axotomy.Methods: Neonatal male mice were used in order to isolate cortical neurons. Isolated primary cortical neurons were cultured. After 24 hours three different rifampicinconcentrations (1 µM, 10 µM and 100 µM) were applied to the neurons and after 15minutes of rifampicin addition, neurons were laser axotomized. Viability of the cellswas evaluated by propidium iodide staining after 24 hours of axotomy.Results: Laser axotomy decreases the cortical neuron viability significantly by80.45%, while rifampicin pre-treatment increases their viability in all three dosages ina statistically significant manner.Conclusion: Rifampicin has an acute neuroprotective effect on the viability of thelaser axotomized cortical neurons.