This study investigated whether herbal cake-separated moxibustion combined with atorvastatin modifies lipid metabolism, aortic pathological structure, plaque deposition, and the integrin/YAP/JNK signaling pathway in ApoE-/- mice with diet-induced atherosclerosis (AS). The aim was to explore potential preventive and therapeutic mechanisms relating to endothelial dysfunction and inflammatory activity in the aorta.
Animals and groups: Male C57BL/6J mice were used as a blank control group. Male ApoE-/- mice were randomized into four groups (n = 9 per group): model (untreated), medicine (atorvastatin), herbal cake-separated moxibustion, and combined treatment (atorvastatin plus moxibustion). A high-fat diet was used to establish the AS model over the experimental period.
Interventions: The herbal cake-separated moxibustion protocol applied moxibustion at the acupoints Danzhong (CV17) and Shenque (CV8), once every other day, three times per week. The medicine group received atorvastatin calcium by gavage at 3 mg·kg-1·d-1, once daily. The combined group received both interventions simultaneously. Treatments continued for 8 weeks.
Outcome measures: Body weight was recorded before and after treatment. Aortic pathological morphology was assessed by hematoxylin-eosin (HE) staining; aortic lipid plaque area was quantified using Oil Red O staining. Serum lipid measures (triglyceride [TG], total cholesterol [TC], LDL-C, HDL-C) were obtained by biochemical analysis. Serum biomarkers of endothelial function and inflammation—vascular endothelial growth factor (VEGF), endothelin-1 (ET-1), interleukin (IL)-6, IL-8, tumor necrosis factor-α (TNF-α)—were measured by ELISA. Serum nitric oxide (NO) was measured by a colorimetric assay. Aortic protein expression of integrin αVβ3, YAP, phosphorylated YAP (p-YAP), phosphorylated JNK1/2 (p-JNK1/2), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) was determined by Western blot.
Statistical reporting: The abstract reports group comparisons with P values (P < 0.05 or P < 0.01) for the described endpoints. Exact statistical tests, variance measures, and raw numerical values beyond directional change were not provided in the abstract.
Model validation: Compared with blank controls, ApoE-/- model mice had significantly increased body weight (P < 0.01) and clear aortic pathology: thickened intima, foam cell and lipid accumulation, smooth muscle cell hypertrophy and edema, thinning of elastic fibers, widened endothelial gaps, and ruptured elastic membranes. Arterial lipid plaque area was significantly larger (P < 0.01). Serum TC, TG, LDL-C, IL-6, IL-8, TNF-α, ET-1, and VEGF were elevated (P < 0.01), while HDL-C and NO were reduced (P < 0.01). Aortic protein expression of integrin αVβ3, YAP, p-JNK1/2, ICAM-1, and VCAM-1 increased, whereas p-YAP decreased (all P < 0.01).
Effects of interventions versus model: The atorvastatin (medicine), moxibustion, and combined treatment groups all showed significant improvements compared to the model group. Body weight decreased in these treatment groups (P < 0.01), and the magnitude of weight change was significantly lower than in the model group. Aortic lumen structure was more regular, intimal thickness was reduced, foam cells and lipid deposition were decreased, and smooth muscle cell edema, hypertrophy, and inflammatory infiltration were alleviated. Most biochemical and inflammatory markers showed significant improvement compared to the model (P < 0.05 or P < 0.01).
Comparisons among treatments: Compared with the moxibustion group, both the medicine and combined groups had smaller plaque area (P < 0.05 or P < 0.01), higher serum NO (P < 0.01), lower LDL-C, ET-1, and VEGF (P < 0.01), and reduced aortic p-JNK1/2, ICAM-1, and VCAM-1 protein expression (P < 0.05 or P < 0.01).
Specific atorvastatin effects: The medicine group showed decreases in serum TC, TG, IL-6, IL-8, and TNF-α (P < 0.01), an increase in HDL-C (P < 0.01), reduced aortic integrin αVβ3 and YAP protein expression (P < 0.01 and P < 0.05 respectively), and increased p-YAP protein expression (P < 0.05).
Combined therapy outcomes: Relative to either monotherapy, the combined treatment group had greater body weight reduction (P < 0.05 or P < 0.01), larger decreases in serum TC, TG, IL-6, IL-8, TNF-α, and reduced aortic integrin αVβ3 and YAP expression (P < 0.01 or P < 0.05). The combined group also had higher serum HDL-C (P < 0.01) and increased aortic p-YAP expression (P < 0.01).
Across comparisons, aortic expression changes in integrin αVβ3, YAP, p-YAP, and p-JNK1/2 tracked with disease and treatment effects. The ApoE-/- model showed elevated integrin αVβ3 and YAP with decreased p-YAP and increased p-JNK1/2, consistent with pathological activation of associated signaling in the aorta. Atorvastatin alone reduced integrin αVβ3 and YAP and increased p-YAP; it also reduced p-JNK1/2 in the aorta. Combined treatment produced larger decreases in integrin αVβ3 and YAP and increased p-YAP versus monotherapy. The authors interpret these changes as evidence that the therapeutic benefits—improved lipid profiles, attenuated endothelial dysfunction, and reduced inflammation—may be mediated at least in part by modulation of the integrin/YAP/JNK signaling axis in the aorta.
The abstract reports that herbal cake-separated moxibustion combined with atorvastatin exerts synergistic effects in ApoE-/- mice with diet-induced atherosclerosis. The combined approach improved serum lipid parameters, mitigated aortic endothelial injury, and reduced inflammatory responses. The data suggest these effects may be related to regulation of the aortic integrin/YAP/JNK signaling pathway. Detailed numeric results, full statistical methods, and safety or adverse-event data are not reported in the abstract and would require review of the full text.
All authors declared no conflict of interest in the source report.