The publisher's final edited version of this article is available at Neurochem Int See other articles in PMC that cite the published article. Female ICR mice were randomly divided into ovariectomized or sham-operated groups, and biochemical analyses were carried out at 5 weeks after the operation. The well-known risk factors for AD are increasing age, atherosclerosis, diabetes mellitus, stroke and ApoE 4 polymorphism.
Moreover, clinical studies have shown that ovarian hormone deprivation after menopause can increase AD prevalence Tang et al. Indeed, Yan et al. On the other hand, RAGE, known as a member of the immunoglobulin superfamily of cell surface molecules, is expressed in a variety of cell types including neurons Brett et al. Recent study further indicated that estrogen decreased brain mitochondrial ROS production Razmara et al.
Materials and methods 2. Thus, in this study, biochemical analyses were performed 5 weeks after the operation. The brains were immediately removed, and the hippocampi were dissected away and lysed in with standard diluent and disrupted with a handheld homogenizer. The diluted sample, standards, AChE tracer and rabbit anti-estradiol were put into the appropriate wells of microtiter plates coated with mouse anti-rabbit IgG.
All wells were further washed five times and reacted with TMB solution. Data are expressed as a percentage of the control. Tissue blocks of the hippocampal CA1 and CA3 regions, and dentate gyrus were isolated from coronal sections on ice under a microscope with fold magnification, homogenized in ice-cold lysis buffer 50 mM Tris, pH 7. The separated proteins were transferred electrophoretically to a hydrophobic polyvinylidene diflouride PVDF membrane Millipore Co.
Membranes were then washed and incubated for 1 h at room temperature with horseradish peroxidase-conjugated anti-mouse IgG 1: To quantify the relative amount of proteins, the blots were incubated with the primary antibody detecting GAPDH clone 6C5, 1: After the reaction, the sections were washed five times with 0. To prevent variability in staining due to each experimental procedure, the brains of four mice, equated across experimental groups, were processed at the same slide using the same reagents and temperature conditions.
Statistical analysis The experimental data were statistically analyzed using StatView 5. The significance of differences was determined by the unpaired t-test.