Clitoria ternatea and Moringa oleifera extracts reduce endothelial activation and intimal thickening in diet-induced atherosclerosis

Atherosclerosis is a chronic inflammatory vascular disease characterized by endothelial activation and progressive thickening of the tunica intima. This study aimed to evaluate the effects of combined Clitoria ternatea and Moringa oleifera (CT-MO) extracts, designated as CT-MO, on endothelial activation markers and tunica intima thickness in male Wistar rats fed a high-fat and high-cholesterol (HFHC) diet. Phytochemical screening and ultraviolet-visible (UV-Vis) spectrophotometry were conducted to determine the optimal extract formulation, which was subsequently administered in vivo at doses of 200, 400, and 600 mg/kg body weight for 45 days. Serum levels of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6), and cluster of differentiation 40 (CD40) were measured using enzyme-linked immunosorbent assay (ELISA), while tunica intima thickness was assessed histologically. Correlation and exploratory multivariate regression analyses were conducted to evaluate associations between inflammatory biomarkers and vascular structural remodeling. The CT-MO (25:75) combination demonstrated the highest flavonoid content (79.10 mg/g). The HFHC diet significantly increased inflammatory biomarkers and tunica intima thickness (12.22 µm). Treatment with CT-MO extract significantly reduced VCAM-1, ICAM-1, IL-6, and CD40 levels in a dose-dependent manner (p < 0.05). Correlation analysis demonstrated significant positive associations between inflammatory biomarkers and tunica intima thickness, while exploratory multivariate analysis identified VCAM-1 as the biomarker most strongly associated with vascular structural remodeling. The highest dose (600 mg/kg) exhibited the most pronounced effect, with biomarker levels not significantly different from healthy controls and tunica intima thickness reduced to 3.42 µm. These findings suggest that the combined CT-MO extract may attenuate endothelial activation and vascular structural remodeling associated with early atherosclerosis.