2.10 Enzyme-linked immunosorbent assay (ELISA)
Rat ELISA kits were used to measure the levels of interleukin-1β (IL-1β) (ABclonal, Cat# RK00009, Wuhan, China) and interleukin-6 (IL-6) (ABclonal, Cat# RK00020, Wuhan, China) in the PrL and IL. Briefly, the tissue was homogenized and centrifuged at 2000×g for 20 minutes at 4°C, then the supernatant was extracted and incubated in a 96-well plate. The cytokine levels were estimated by interpolation from a standard curve by colorimetric measurements at 450 nm (correction wavelength 630 nm) on an ELISA plate reader (Thermo Fisher Scientific Multiskan Mk3, MA, USA). The results are shown as pg mg-1 of protein.