Protective Effects of Berberine-Loaded Black Seed (<i>Nigella sativa</i> L.) Oil Nanoemulsion in the Atherosclerotic Mouse Model

Abstract

Background: This study investigated atherosclerosis (AS), a pivotal contributor to the onset of coronary artery disease and other cardiovascular ailments.‎ Objectives: The presents study examined a novel therapeutic strategy combining black seed oil [Nigella sativa L. (NS)] and berberine (BBR) in a C57BL/6J mouse model of AS. Methods: After preparing the NS nanoemulsion, gas chromatography-mass spectrometry (GC-MS) was used for analysis. A BBR-loaded black seed oil-based nanoemulsion (BBS) was developed and evaluated for its thermodynamic stability, viscosity, particle size, and dynamic light scattering (DLS) properties. Forty-eight male C57BL/6J mice (8 weeks old, 18 - 20 g) were divided into six groups and fed a modified AIN-76 diet with 25% dietary fat for 16 weeks to induce AS. Treatment began after 8 weeks through oral gavage for the remaining 8 weeks. Levels of low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride (TG), and total cholesterol were measured, along with the atherogenic coefficient and cardiac risk ratio. We also quantified malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) activities, and assessed pathological changes in cardiac and aortic tissues using hematoxylin and eosin (H&E) staining. Results: The GC-MS analysis confirmed that NS oil met quality benchmarks, displaying stability with a zeta potential of -18.4 mV. The study showed a Polydispersity Index (PDI) of 0.3019 and a Z-average of 250.4 nm. The BBS improved lipid profiles in mice, increasing HDL and decreasing LDL, TG, and total cholesterol, thereby reducing cardiac risk. The formulation exhibited strong antioxidant effects, with reduced MDA levels and enhanced SOD, GPX, and CAT activities. Pathological observations supported these biochemical results. Conclusions: Our findings suggest that BBS offers valuable insights into the mechanisms underlying atherosclerotic disease. This understanding could pave the way for novel approaches to cardiovascular health and the development of effective preventive strategies.