Background: It is now well established that perturbations in cholesterol levels are crucial in the onset and development of Alzheimer’s disease (AD). Despite this evidence, scarce information is available about prospective alterations of proteins controlling brain cholesterol homeostasis in Alzheimer’s disease. Thus, the aim of this work was to analyze the protein expression of enzymes and proteins involved in brain cholesterol biosynthesis and uptake in a mouse model of AD. Methods: Transgenic Tg2576 mouse was used as experimental model of Alzheimer’s disease. Phosphorylated HMGCR, AMPK, LDLr, LRP1 and SR-B1 protein levels were assessed by Western blot in both hippocampus and brain cortex at different disease stages. Results: HMGCR phosphorylation, AMPK and LDLr levels were significantly increased in the hippocampus of 9- and 15-month-old Tg2576 mice. At 15 months of age, hippocampal LRP1 and SR-B1 were also upregulated. On the contrary, no differences were detectable in the analyzed proteins in the brain cortex, at both stages. Conclusion: Marked alterations of proteins regulating cholesterol homeostasis were found in the hippocampus of Tg2576, further sustaining the crucial involvement of cholesterol metabolism in AD.