Ba3677

H&E staining was used to observe the neuronal morphology and damage, while immunohistochemistry was performed to detect the activation of NLRP3 and microglia in the rat hippocampus. The brain tissues were embedded in paraffin, and 5-μm thick brain sections were prepared using a slicer (RM2235, Leica, Germany). After dewaxed and rehydrated by xylene and ethanol, the slides were stained with H&E staining or immunohistochemistry. H&E staining was conducted on 5 μm sections using standard H&E staining protocol. For immunohistochemistry, the sections were subjected to induced antigen retrieval in citrate buffer at 100°C (0.01 M, pH = 6.0), and then immersed in 3% hydrogen peroxide solution for 10 min at RT to quench endogenous peroxidase activity. After blocking with 5% bull serum albumin, the tissues were incubated with anti-NLRP3 antibody (1:50, BA3677, Boster, Wuhan, China) or anti-Iba-1 antibody (1:500, ab178846, Abcam, USA) overnight at 4°C. Afterward, the sections were rinsed with PBS three times and incubated with anti-rabbit IgG secondary antibodies (1:200, Boster, Wuhan, China) at RT for 1 h, and then were viewed by 3,3′-diaminobenzidine solution and counterstained with Hematoxylin. The sections were visualized and analyzed by a light microscope (Olympus, Tokyo, Japan) equipped with an imaging system and Image J software.