Atherosclerosis (AS) is the common pathological basis of chronic cardiovascular diseases and is associated with inflammation and lipid metabolism dysfunction. Geniposide, the main active ingredient of Gardenia jasminoides Ellis fruit, exhibits a variety of anti-inflammatory and anti-oxidative functions; however, its role in AS remains unclear. The aim of this study was to investigate the mechanisms of geniposide in alleviating inflammation and thereby attenuating the development of AS. ApoE-/- mice were fed a high fat diet to induce AS and were treated with geniposide (50 mg/kg) for 12 weeks. Blood glucose and lipid levels were measured by biochemical analysis. H&E, Masson and Oil red O staining were performed to observe morphological changes and lipid deposition in the aorta and liver. Serum inflammatory cytokines were detected by ELISA. Dual-luciferase reporter gene assay was used to verify the target relationship between microRNA-101 (miR-101) and mitogen-activated protein kinase phosphatase-1 (MKP-1). The levels of miR-101, p-p38, and MKP-1 in the aorta were detected by qPCR and western blotting. The anti-inflammatory effect of geniposide in vitro was investigated in the RAW264.7 macrophage cell line. A miR-101 mimic and an inhibitor were used to study the effect of miR-101 on regulating the expression of the target MKP-1 and the downstream inflammatory cytokines. Geniposide treatment reduced lipid levels and plaque size in the mouse model of AS. Geniposide downregulated miR-101 to upregulate MKP-1 and suppress the production of inflammatory factors in vitro and in vivo. Geniposide suppressed the levels of inflammatory factors in the presence of the miR-101 mimic, whereas no obvious effect was observed in the miR-101 inhibitor group. We concluded that geniposide reduced the plaque size and alleviated inflammatory injury in ApoE-/- mice and RAW264.7 cells. The specific anti-inflammatory mechanism was related to the miR-101/ MKP-1/p38 signaling pathway.

Gardenia jasminoides Ellis is a traditional Chinese medicine that has been used for treatment of various diseases, including atherosclerosis by clearing heat and detoxication. Geniposide is considered as the effective compounds responsible for the therapeutic efficacy of Gardenia jasminoides Ellis against atherosclerosis. To investigate the effect of geniposide on atherosclerosis burden and plaque macrophage polarization, with focus on its potential impact on CXCL14 expression by perivascular adipose tissue (PVAT). ApoE^-/- mice fed a western diet (WD) were used to model atherosclerosis. In vitro cultures of mouse 3T3-L1 preadipocytes and RAW264.7 macrophages were used for molecular assays. The results revealed that geniposide treatment reduced atherosclerotic lesions in ApoE^-/- mice, and this effect was correlated with increased M2 and decreased M1 polarization of plaque macrophages. Of note, geniposide increased the expression of CXCL14 in PVAT, and both the anti-atherosclerotic effect of geniposide, as well as its regulatory influence on macrophage polarization, were abrogated upon in vivo CXCL14 knockdown. In line with these findings, exposure to conditioned medium from geniposide-treated 3T3-L1 adipocytes (or to recombinant CXCL14 protein) enhanced M2 polarization in interleukin-4 (IL-4) treated RAW264.7 macrophages, and this effect was negated after CXCL14 silencing in 3T3-L1 cells. In summary, our findings suggest that geniposide protects ApoE^-/- mice against WD-induced atherosclerosis by inducing M2 polarization of plaque macrophages via enhanced expression of CXCL14 in PVAT. These data provide novel insights into PVAT paracrine function in atherosclerosis and reaffirm geniposide as a therapeutic drug candidate for atherosclerosis treatment.

Dingxin Recipe III (DXRIII) is a traditional Chinese medicinal formulation that has been employed in clinics for over two decades. It is utilized in the treatment of cardiovascular diseases associated with atherosclerosis (AS) through mechanisms purported to involve the clearing of heat and detoxification, as well as the promotion of blood circulation and the removal of blood stasis. Despite its widespread application and reported therapeutic benefits, its exact mechanisms remain incompletely elucidated. This study aimed to confirm the anti-AS efficacy of DXRIII and elucidate its underlying mechanism, specifically focusing on the amelioration of endothelial senescence. The mechanisms underlying the anti-AS effects of DXRIII were elucidated through network pharmacology analysis. In vivo and in vitro models were established using high-fat diet (HFD) induced ApoE^-/- mice and H₂O₂-induced human umbilical vein endothelial cells (HUVECs), respectively. The histopathologic changes of aortic tissues, inflammatory response, oxidative stress, mitochondrial damage, mitophagy, senescence, and FOXO3a/PINK1/Parkin signaling pathway were evaluated through histological staining, transmission electron microscopy, fluorescent probe staining, RT-qPCR, and Western blot. The constituents of DXRIII were identified via liquid chromatography-mass spectrometry, and the binding affinity and stability of its active compounds with target proteins were investigated using molecular docking and cellular thermal shift assay. Foxo3a was silenced in vitro by lentiviral transfection to determine its involvement in DXRIII-induced mitophagy. DXRIII effectively ameliorated plaque pathology in AS mice and attenuated endothelial cell senescence. Meanwhile, DXRIII suppressed pro-inflammatory factors and adhesion molecules, mitigated oxidative stress and mitochondrial damage, while activating PINK1/Parkin-mediated mitophagy and upregulating FOXO3a expression. Notably, both mitophagy inhibition Mdivi-1 and silencing of FOXO3a in vitro blocked DXRIII's anti-senescence effects. Furthermore, the active ingredients of DXRIII, including berberine, kaempferol, quercetin and luteolin, showed strong binding affinity with FOXO3a and enhanced its protein stability. Our findings for the first time demonstrated that DXRIII effectively alleviates endothelial senescence and HFD-induced AS, possibly by activating FOXO3a and subsequently enhancing PINK1/Parkin-mediated mitophagy.

Background: The prognostic value of discharge renin–angiotensin–aldosterone system inhibitor (RASi) therapy in contemporary PCI-treated acute myocardial infarction (AMI) survivors with preserved or mildly reduced left ventricular ejection fraction (LVEF) remains uncertain. Methods: A retrospective cohort study of 2530 AMI patients (2019–2022) stratified by RASi use. Exclusion criteria were in-hospital mortality, LVEF < 40%, contraindications to the use of RASis or no percutaneous coronary intervention (PCI). Primary endpoints included heart failure (HF) events, recurrent acute coronary syndrome (ACS), and all-cause mortality. Kaplan–Meier analyses and inverse probability of treatment weighting (IPTW)-weighted Cox models were applied. Results: Over a mean follow-up of 49 months, discharge RASi therapy was not associated with all-cause mortality overall, but was associated with fewer HF rehospitalizations (HR 0.62, 95% CI 0.40–0.95; p = 0.03). Mortality associations differed by AMI type and hypertension status, particularly for NSTEMI (HR 0.36, 95% CI 0.14–0.91; p = 0.03; p for interaction = 0.02) and hypertension (HR 0.36, 95% CI 0.15–0.84; p = 0.02; p for interaction = 0.04). Conclusions: In this single-center observational cohort of PCI-treated AMI survivors with LVEF ≥ 40%, discharge RASi therapy was associated with fewer HF rehospitalizations but not with lower overall mortality. Exploratory subgroup analyses suggested potential heterogeneity according to NSTEMI status and hypertension, but these findings should be considered hypothesis-generating and require confirmation.