This study was conducted to evaluate the possible protective effect of Spirulina and alpha-lipoic acid against aspartame induced oxidative stress and brain damage in rabbits. Forty-two white male New-Zealand Rabbits were classified into seven groups. Group I(Control) received no drugs. Group II (ASP) received aspartame (250 mg/kg b. wt./day). Group III (LA)received alpha-lipoic acid (100 mg/kg b. wt./day). Group IV (SPir) received spirulina platensis (1500 mg/kg. b. wt./day). Group V (ASP+ LA) received aspartame (250 mg /kg b. wt.) and alpha-lipoic acid (100 mg/kg b. wt.). Group VI (ASP+ Spir) received aspartame (250 mg/kg b.wt.) and spirulina platensis (1500 mg/kg b. wt.). Group VII (ASP+ALP+Spir) received aspartame (250 mg/kg b.wt.), alpha-lipoic acid (100 mg/kg b. wt.) and Spirulina platensis (1500 mg/kg b. wt.) for 8 weeks. At the end of experiment, brain tissues were collected and analyzed for L-malondialdehyde(L-MDA), superoxide dismutase (SOD), reduced glutathione (GSH), gene expression of tumor necrosis factor alpha (TNF-α)and caspase 3, and DNA damage. The results showed a significant up-regulation of TNF-α and caspase 3 gene expression level, marked increase in L-MDA and DNA damage, and a marked decrease in SOD activity and GSH concentration in ASP groupwhen compared with control group. Co-administration of spirulina and alpha-lipoic acid protected aspartame induced brain damage and caused a significant improvement of all previous parameters and attenuated DNA changes. Conclusively, Spirulina platensis and alpha-lipoic acid exerted a protective effect against DNA damage and oxidative stress in the brain of aspartame usage through free radical scavenging and anti-inflammatory activities as well as regenerating endogenous antioxidants defense system mechanisms.