Ferric chloride-induced murine carotid arterial injury: A model of redox pathology

Abstract

Ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for platelet aggregation to occlude blood flow gives a quantitative measure of vascular damage that is pathologically relevant to thrombotic disease. We have refined the traditional FeCl3-induced carotid artery model making the data highly reproducible with lower variation. This paper will describe our artifices and report the role of varying the oxidative damage by varying FeCl3 concentrations and exposure. To explore a maximum difference between experimental groups, adjustment of the selected FeCl3 dose and exposure duration may be necessary.

Keywords: Animal model; CA, carotid artery; Carotid artery; Ferric chloride; Thrombosis.

Graphical abstract

Fig. 1

Procedures to expose the jugular vein, injection of rhodamine 6G flurescence dye into the circulation system and exposure of left carotid artery. T indicates trachea. See text for details.

Fig. 2

Effect of different concentration of FeCl₃ on thrombosis. Representative video images of thrombi formation in carotid artery (CA) treated with different concentration of FeCl₃, 2.5% (A), 5% (B), 7.5% (C), and 10% (D) are shown. Blood flow was from right to left. “Before” indicates images taken immediately prior to FeCl₃ treatment. Numbers indicate time in minutes after removing the filter paper. Arrows in A indicate the tiny thrombi which were first observed.

Fig. 3

Vessel occlusion times after different concentrations of FeCl₃ treatment. Mice were treated as in Fig. 2 and occlusion time assessed by intravital microscopy. *p<0.01 wt vs. cd36 null at the same concentration of FeCl₃. One-way ANOVA Bonferroni/Dunn was used to determine the differences among groups. Data are represented as mean±SEM. N=3 for 2.5% FeCl₃ treated wt and cd36 null mice; n=4 for 5% FeCl₃ treated cd36 null mice; and n=5 in all other conditions.

Fig. 4

Duration of FeCl₃ treatment on thrombosis. (A) Representative video images of thrombi in CA exposed to 5% FeCl₃ for 1, 3 or 5 min. “Before” indicates before FeCl₃ treatment. Numbers indicate time in minutes after removing the filter paper. (B) Statistical analysis of vessel occlusion time. N=3 for 1 min and n=5 for 3 and 5 min treatments. One-way ANOVA (Bonferroni/Dunn) was used to determine the differences among groups. Data are represented as mean±SEM.