2.10 Enzyme-linked immunosorbent assay (ELISA)
After the completion of behavioral tests, rats were deeply anesthetized with pentobarbital sodium (30 mg/kg) and then sacrificed by decapitation. Then, BLA tissue was collected and prepared using a grinder and an ultrasonic tissue homogenizer. Tumor necrosis factor (TNF)-α (KE20001, ProteinTech Group, Chicago, IL, USA), interleukin (IL)-1β (KE20005, ProteinTech Group, Chicago, IL, USA), CXCL1 (EK0724, Boster Biological Technology, Pleasanton, CA, USA) and MCP-1 (EK0902, Boster Biological Technology, Pleasanton, CA, USA) were detected using ELISA kits in accordance with the manufacturer’s instructions. The optical density (OD) values were assessed with a microplate reader (NK3; Ladsystems, Helsinki, Finland) at 450 nm, and the concentrations of TNF-α, IL-1β, CXCL1 and MCP-1 were calculated according to the standard curve and presented as pg/mg protein in the tissue homogenate.