Nanoparticle (NP) forms of aluminium oxide (Al2O3) are used in various fields such as engineering, pharmacy, medicine etc. Compounds containing aluminium oxide NPs may present toxic effects after certain thresholds. Thus, the present study was carried out to determine the effects of Al2O3 nanoparticles (Al-NPs) in rats. For this aim, different doses (0, 0.5, 5, 50 mg/kg b.w./day) of Al NP ((similar to)40 nm) were orally administered to female rats (Rattus norvegicus var. albinus) for 14 days and the response of several biomarkers such as activities of ATPases (total ATPase, Na,K-ATPase, Mg-ATPase) and acetylcholinesterase (AChE), levels of different glutathione forms and thiobarbituric acid reactive substances (TEARS) were measured in different tissues. Additionally, tissue accumulation of Al-NPs was demonstrated by a transmission electron microscope (TEM). The images showed the presence of Al-NP aggregates in all the tissues at all doses. The sizes of NP aggregates were dependent on NP doses and it was a bit more loose in the brain than in the liver and kidney. AChE activity in the brain decreased significantly at all NP doses, whereas TEARS levels in the liver did not alter significantly at any NP dose. Although there was no significant change in ATPase activities in the intestine at any NP dose, there were significant decreases in the kidney and brain. There were some variations in the levels of total glutathione (tGSH), oxidized glutathione (GSSG) and reduced glutathione (rGSH), though these variations were not significant (P > 0.05). Likewise, the ratio of rGSH/GSSG also did not differ significantly among NP doses and control. The brain seems most affected organ following Al-NP administration. This study demonstrated that most biomarkers in the tissues of rats were affected by Al-NP, showing the signal of toxic effects and suggests further studies to understand better the effects of Al NPs, especially in their use for pharmacology.