Protective effects of Yiqi Huoxue Huatan formula on atherosclerosis: An in vivo and in vitro study.

Atherosclerosis (AS) is a significant pathological contributor to cardiovascular disease, marked by high rates of death and illness globally. Yiqi Huoxue Huatan formula (YHH), a Chinese traditional decoction, has been clinically validated as effective for patients with AS; however, its underlying mechanisms remain partially unexplained. The goal of this research is to understand the protective effects of YHH against AS and the mechanisms involved. In vivo, YHH was administered orally at 14.24 and 28.47 g/kg/day (raw medicinal herbs) to ApoE mice on a high-fat diet (HFD) to model AS. In vitro, RAW264.7 macrophages were exposed to oxidized low-density lipoprotein (ox-LDL) and YHH-conducting serum. The therapeutic effects of YHH were assessed using biochemical assays, histopathological observations, and enzyme-linked immunosorbent assays. Mass spectrometry, network pharmacology analyses, molecular docking, and western blotting were used to identify the primary components and anti-AS mechanisms of YHH. YHH significantly reduced AS development, as evidenced by decreased aortic plaque and lower blood lipid levels in HFD-induced AS mice. Meanwhile, YHH significantly decreased the increase in inflammatory cytokines in the supernatant of macrophages induced by ox-LDL. Mechanistic studies revealed that YHH significantly downregulated protein levels of TLR4, p-NF-κB, and HIF-1α. Moreover, molecular docking showed that key active compounds in YHH, including Gardenin A, isosinensetin, alisol B, 3',4',5',5,7-pentamethoxyflavanone, and scutellarein, exhibited strong binding activity to the core targets. YHH emerges as a promising agent in ameliorating AS by reducing inflammatory responses, achieved via the downregulation of TLR4/NF-κB/HIF-1α signaling pathways.