2.7. Enzyme-linked immunosorbent assay (ELISA)

Blood samples were taken from 15 rats, which were decapitated under isoflurane and pentobarbital anesthesia the day after surgery. The blood samples were anticoagulated with ethylenediaminetetraacetic acid and centrifuged at 1000 G. The isolated hippocampus was homogenized in neuronal protein extraction reagent buffer (Thermo Fisher Scientific, Waltham, MA, USA) containing protease inhibitor (ThemoFisher Scientific). After homogenization, the lysates were centrifuged at 100 G. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels were determined using ELISA kits specific for rat TNF-α and IL-1β (R&D Systems, Inc., Minneapolis, MA, USA) according to the manufacturer's instructions. The absorbance values were measured at 450 nm and 540 nm using a micrometer plate reader.

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