Effect of Inhibition of Galectin-3 Expression on Lipid Accumulation and Atherosclerosis Lesions in a Rat Model of Carotid Atherosclerosis

Abstract

Objective: Galectin-3 (Gal-3), is a crucial protein involved in regulating cell adhesion, inflammation, and fibrosis, thereby playing a pivotal role in the occurrence and progression of Atherosclerosis (AS). Using the rat carotid artery AS model, this study aimed to investigate the regulatory effects of Galectin-3 expression on lipid accumulation and AS lesion. Methods: Thirty male Wistar rats, aged 10 weeks, were randomly assigned to three groups: the control, model, and treatment groups, each comprising 10 rats. In the control group, dissection was limited to the left common carotid artery. Moreover, the carotid atherosclerosis model was induced in both the model and treatment groups. The treatment group was administered with 10% Modified citrus pectin (MCP) concurrently one week before surgery. Blood and tissue samples were collected on the day after the operation, and the atherosclerosis modeling was observed using Hematoxylin-Eosin (H&E) staining. The levels of triglyceride, total cholesterol, low-density lipoprotein, and high-density lipoprotein were assessed in these three rat groups. Oil red O staining was used to analyze the lipid deposition in the intima of arteries. Furthermore, the expression levels of Cholesterol ester hydrolase (CEH) and Gal-3 in the carotid artery were determined by quantitative Real-time Polymerase Chain Reaction (qRT-PCR) and Western blot analysis. Enzyme-Linked immunosorbent assay (ELISA) was employed to assess the expression levels of Gal-3, tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF), Interleukin 6 (IL-6), and Interleukin 8 (IL-8). Additionally, flow cytometry was utilized to evaluate mitochondrial membrane potential (MMP) and the expression of iron death-related proteins (Glutathione Peroxidase 4 and Nicotinamide Adenine Dinucleotide Phosphate (NADPH) oxidase 4 (NOX4)) was determined using Western blot analysis. Results: Gal-3 was highly expressed in atherosclerotic rats. After the inhibition of Gal-3, a significant reduction was observed in the levels of triglyceride, total cholesterol, and low-density lipoprotein cholesterol in tissues. The degree of lesions, the levels of inflammatory factors and iron death were significantly alleviated, while the expression of CEH was increased. Conclusion: Inhibition of Galectin-3 can slow down the lesion and lipid accumulation in carotid atherosclerotic rats.