E el r0012

The serum cytokines; tumor necrosis factor-alpha (TNF-α), interferon gamma (IFN-γ) and interleukin-1 beta (IL-1β), were measured by specific ELISA kits (Elabscience, USA catalog numbers E-EL-R2856, E-EL-R0009, and E-EL-R0012, respectively) based on quantitative sandwich technique. The optical density values were read in a micro plate reader at 450 nm. The concentrations of cytokines in samples were determined by comparing the OD of the samples to the standard curve of each measured cytokine. The results were expressed as pg/mg.

The animals were decapitated 24 h after the final AD examination, and the trunk blood was collected in heparinized tubes and centrifuged (2500×g, 15 min, 4 °C). The plasma was aliquoted and stored at − 80 °C until further analysis. After thawing, an extra centrifugation step was applied to prevent nonspecific signals from activated platelets (10,000×g, 10 min, 4 °C). The concentrations of IL-1β, IL-6, UCH-L1, GFAP and MMP-9 were measured using commercially available sandwich-based ELISA kits for rats (Elabscience, E-El-R0012 for IL-1β, detection range of 31.25–2000 pg/ml; Elabscience, E-El-R0015 for IL-6, detection range of 62.5–4000 pg/ml; R&D Systems, RMP900 for MMP-9, detection range of 0.156–10 ng/ml; MyBioSource Inc., MBS 2886354 for GFAP, detection range of 31.2–2000 pg/ml; Elabscience, for UCH-L1, detection range 61.25–4000 pg/ml). The optical density was determined using a microplate reader (SYNERGY H1, BioTek) set to 450 nm.