Ferric Chloride-Induced Carotid Thrombosis

Carotid artery thrombosis was produced using ferric chloride (FeCl₃), as described previously (Ren et al., 2017 (link)). Male adult mice were anesthetized by intraperitoneal (IP) injection of sodium pentobarbital (50 mg/kg) and euthanized by rapid cervical dislocation. Linagliptin (ApexBio, United States) prepared in 0.5% hydroxyethylcellulose and was dosed by gavage (5, 10, or 20 mg/kg/day; once daily for 7 days) (Darsalia et al., 2013 (link); Zhuge et al., 2016 (link)), and vehicle-treated mice were included. The common carotid arteries were exposed, and filter paper (3 × 1.0 mm) soaked in FeCl₃ solution (10%) was placed on top of the left carotid artery for 3 min. After the filter paper was removed, the carotid artery was washed in PBS, and the blood flow was continuously monitored with a vascular flow probe (Transonic Systems, Ithaca, NY, United States) from the onset of injury until stable occlusion occurred (defined as no flow for 120 min).