An Atherosclerosis Model in Mice Induced by Both Partial Carotid Ligation and Overexpression of PCSK9

The lack of rapid, reproducible carotid-specific atherosclerosis models in wild-type mice limits translational studies of plaque biology and therapies. This protocol describes a non-genomically edited, time-efficient method to induce reproducible carotid atherosclerosis in C57BL/6 mice by combining adeno-associated virus (AAV)-mediated PCSK9 overexpression, an atherogenic high-fat diet, and left partial carotid ligation (PCL). The objective is to provide a standardized workflow that reproduces key pathophysiological features of human carotid disease-hyperlipidemia plus disturbed flow-without requiring genetically modified animals. Methods include PCSK9-AAV dosing and administration, perioperative preparation, step-by-step surgical ligation of the left carotid, postoperative care, dietary regimen, tissue collection, and histological assessment including Oil Red O and Verhoeff-Van Gieson staining. The results indicate elevated circulating cholesterol and low-density lipoprotein levels compared with reference values. Pronounced atherosclerotic plaques develop at the left ligated carotid artery, with significantly increased lipid accumulation and significant arterial intima-media thickening relative to the right non-ligated control artery. This standardized protocol improves reproducibility and accessibility for investigators studying carotid plaque formation, progression, intervention strategies, and facilitates cross-laboratory comparisons of experimental therapies.