Mouse model of carotid artery ligation

The carotid artery ligation (CAL) was performed according to the protocol developed by Allagnat et al (14), on 8-week-old male C57BL/6J mice, as previously described. Mice were fed standard laboratory chow and water ad libitum, and were housed in single cages under specific pathogen-free conditions (temperature and humidity were controlled at 22±2°C and 55±5%) in a room with a 12-h light/dark cycle. Male mice (weight, 20–25 g; age, 8 weeks) were used in the present study. Briefly, the mice were anesthetized with 4% chloral hydrate. The left common carotid artery was dissected through a small cervicotomy and ligated with prolene 7.0 just below the carotid bifurcation. The mice were sacrificed by cervical dislocation, and perfused with PBS followed by buffered paraformaldehyde 4% through the left ventricle prior to CAL for reference or on days 3, 14 or 28 days following CAL. All animal experimentation conformed to the Guidelines of the local Animal Care and Use Committee at Tangshan Workers Hospital.