Ziziphora clinopodioides flavonoids based on network pharmacology attenuates atherosclerosis in rats induced by high-fat emulsion combined with vitamin D3 by down-regulating VEGF/AKT/NF-κB signaling pathway

Ziziphora clinopodioides flavonoids (ZCF) is a major bioactive total flavonoids compound isolated from Ziziphora clinopodioides Lam. It has been long used as an anti-atherosclerosis (AS) in clinics. However, anti-AS effects of ZCF have not been fully investigated. The objective of this study is to further investigate the anti-AS activities and mechanisms of ZCF in vivo. The main chemical components, action targets and signal pathways of Ziziphora clinopodioides Lam were predicted and analyzed by network pharmacology technology. The main bioactive components of Ziziphora clinopodioides Lam were identified using high performance liquid chromatography-mass spectrometry (HPLC-MS). In vivo experiments, atherosclerosis in rats induced by high-fat emulsion combined with vitamin D-3 and treated with simvastatin (0.45 mg/kg/d), ZCF (6.25, 12.5, 25 g/kg/d) for 7 weeks. We found that ZCF significantly reduced blood lipid levels (TG, TC, and LDL-C), and decreased lipid deposition in the aorta and atherosclerotic lesion size, inhibited mitochondrial membrane potential (MMP2/9/12/13) impairment. Meanwhile, ZCF may down-regulated the levels of VEGF, AKT, NF-kappa B, ICAM-1 and VCAM-1 proteins, indicating ZCF may play an anti-atherosclerotic role by down-regulating the VEGF/AKT/NF-kappa B signaling pathway. Results from this study demonstrated that ZCF have an anti-AS ability to lower lipid concentrations and protect endothelial function, antioxidant and anti-inflammatory activity, and suggested that ZCF might be a potential therapeutic drug in the prevention of AS.