Background: A failure of the Na+,K+-ATPase activity (which is essential for ion flux across the cell membranes) occurs in many pathological conditions and may lead to cell dysfunction or even cell death. By altering the concentration of specific opioid peptides, gamma-hydroxybutyric acid (GHB) may change ion flux across cell membranes and produce the ‘channel arrest’ which we assumed will inhibit the failure of Na+,K+-ATPase activity and therefore lead to energy conservation and cell protection. Therefore we planned this study to see the effects of GHB at two different doses on Na+,K+-ATPase activity in an experimental head trauma model. Methods: Forty New Zealand rabbits were divided equally into four groups: group I was the sham-operated group, group II (untreated group), group III received head trauma and intravenous (i.v.) 500 mg/kg GHB and group IV received head trauma and i.v. 50 mg/kg GHB. Head trauma was delivered by performing a craniectomy over the right hemisphere and dropping a weight of 10 g from a height of 80 cm. The non-traumatized (left) side was named as ‘a’ and the traumatized (right) side as ‘b’. One hour after the trauma in groups II and III and craniotomy in group I, brain cortices were resected from both sides and in group I only from the right side was the tissue Na-K-ATPase activity determined. Results: The mean ± SD of Na+,K+-ATPase levels of each group are as follows: group I – 5.97 ± 0.55; group IIa – 3.90 ± 1.08; group IIb – 3.58 ± 0.90; group IIIa – 5.53 ± 0.60; group IIIb – 5.33 ± 0.88; group IVa – 5.05 ± 0.72; group IVb – 4.93 ± 0.67. The Na+,K+-ATPase levels of group IIa, IIb, IVa and IVb were significantly different from group S (P < 0.05). There were also significant differences between group IIa and groups IIIa and IVa; group IIb and groups IIIb and IVb (P < 0.05). Conclusions: We conclude that GHB is effective in suppressing the decrease in Na+,K+-ATPase levels in brain tissue at two different dose schedules after head trauma.