Western blotting
The ipsilateral cerebral hemisphere, rat leukocytes, and cultured cells were homogenised in lysis buffer containing phosphatase and protease inhibitors. We used the following primary antibodies: anti‐Interleukin (IL)-1β (Catalogue, AF-501-NA), anti-tumour necrosis factor (TNF)-α (Abcam, ab205587), anti‐Matrix Metallopeptidase(MMP)-9 (Abcam, ab76003), anti‐zonula occludens(ZO)-1 (arigo, ARG55738), anti-occludin (Abcam, ab216327), anti‐caspase-3 (Abcam, ab13847; Affinity Biosciences, AF7022), anti‐C1QTNF6 (Abcam, ab36900), and anti‐β‐actin (Abcam, ab20272). Signals were detected through incubation with horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology) for 60 min at room temperature and using an enhanced chemiluminescence kit (Millipore, Darmstadt, Germany). Grey values of the protein bands were analysed using ALPHAEASE FC software (Alpha Innotech, San Leandro, CA, USA).