To investigate the effects of herbal cake-separated moxibustion combined with atorvastatin on lipid metabolism, aortic pathological structure, plaque deposition, and integrin/yes-associated protein (YAP)/c-Jun N-terminal kinase (JNK) signaling pathway in ApoE^-/- atherosclerosis (AS) mice, and to explore its preventive and therapeutic mechanisms for AS.

Male C57BL/6J mice were used as the blank group, and male ApoE^-/- mice were randomly divided into the model group, medicine group, herbal cake-separated moxibustion group, and combined treatment group, with 9 mice per group. High-fat diet was used to establish the AS model. The herbal cake-separated moxibustion group received moxibustion at “Danzhong” (CV17) and “Shenque” (CV8) acupoints, once every other day, 3 times a week. The medicine group received atorvastatin calcium tablet via gavage, 3 mg·kg^-1·d^-1, once daily. The combined treatment group received an integrated approach that combined the interventions of the medicine group and herbal cake-separated moxibustion group. All groups were treated for 8 weeks. Body weight were observed before and after treatment. HE staining was used to observe aortic pathological morphology, and Oil Red O staining to observe aortic lipid plaque area. Biochemical analysis was used to detect serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels. Serum vascular endothelial growth factor (VEGF), endothelin-1 (ET-1), interleukin (IL)-6, IL-8, and tumor necrosis factor-α (TNF-α) levels were detected by ELISA, and serum nitric oxide (NO) level was detected by colorimetric assay. Western blot was used to detect protein expression levels of integrin αVβ3, YAP, phosphorylated (p)-YAP, p-JNK1/2, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in the aorta.

Compared to the blank group, the model group exhibited increased body weight (P<0.01); thickened aortic intima, foam cells and lipid deposition, hypertrophic edema of smooth muscle cells, thinning of elastic fibers, widened endothelial cell gaps, and rupture of elastic membranes; significantly increased arterial lipid plaque area (P<0.01); elevated serum TC, TG, LDL-C, IL-6, IL-8, TNF-α, ET-1, and VEGF levels (P<0.01), while HDL-C and NO levels decreased (P<0.01); increased protein expression of integrin αVβ3, YAP, p-JNK1/2, ICAM-1, and VCAM-1 (P<0.01), and reduced p-YAP protein expression (P<0.01) in the aorta. Compared to the model group, the medicine group, herbal cake-separated moxibustion group, and combined treatment group showed reduced body weight (P<0.01), with a significantly lower weight change difference than that of the model group (P<0.01); more regular aortic lumen structure, thinner intima, reduced foam cells and lipid deposition, alleviated smooth muscle cell edema, hypertrophy, and inflammatory infiltration; all other indicators also showed significant improvement compared to the model group (P<0.01, P<0.05). Compared to the herbal cake-separated moxibustion group, the medicine group and combined treatment group exhibited reduced plaque area (P<0.05, P<0.01), increased serum NO level (P<0.01), decreased LDL-C, ET-1, and VEGF levels (P<0.01), and reduced protein expressions of p-JNK1/2, ICAM-1, and VCAM-1 in the aorta (P<0.05, P<0.01); the medicine group also showed decreased serum TC, TG, IL-6, IL-8, and TNF-α levels (P<0.01), increased HDL-C level (P<0.01), reduced integrin αVβ3 and YAP protein expressions in the aorta (P<0.01, P<0.05), and increased p-YAP protein expression (P<0.05). Compared to the medicine group and the herbal cake-separated moxibustion group, the combined treatment group exhibited a greater reduction in body weight (P<0.05, P<0.01), decreased serum levels of TC, TG, IL-6, IL-8, and TNF-α, as well as reduced expressions of integrin αVβ3 and YAP proteins in the aorta (P<0.01, P<0.05), elevated serum HDL-C level (P<0.01), and increased p-YAP protein expression (P<0.01).

Herbal cake-separated moxibustion combined with atorvastatin exerts synergistic effects, ameliorates serum lipid profiles, mitigates endothelial dysfunction of aorta, and attenuates inflammatory responses in AS mice, potentially via modulation of the integrin/YAP/JNK signaling pathway in the aorta.

To observe the clinical effect and safety of herbal cake-insulated moxibustion in the auxiliary treatment of carotid atherosclerosis (CAS) differentiated as interaction of phlegm and blood stasis.

Sixty-three patients with CAS differentiated as interaction of phlegm and blood stasis were randomly divided into an experimental group (33 cases, 4 cases dropped out) and a control group (30 cases, 3 cases dropped out). In the experimental group, herbal cake-insulated moxibustion was delivered in combination with oral administration with atorvastatin. In the control group, atorvastatin was administered orally. The duration of treatment in either group was composed of 12 weeks. Common carotid intima-media thickness (cIMT), the total area of carotid plaque, common carotid blood flow parameters, blood lipids contents (4 items) and traditional Chinese medicine (TCM) syndrome score were compared between the two groups before and after treatment. Blood routine and liver and kidney function were detected before and after treatment, and adverse events were recorded to evaluate safety.

Compared with those before treatment, the bilateral cIMT and the contents of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in serum were reduced after treatment (P<0.05, P<0.001), the scores of the primary symptoms and secondary symptoms of TCM, as well as the total score were lower (P<0.001) in the two groups. The left cIMT and the contents of the serum TC, TG and LDL-C in the experimental group were lower than those of the control group (P<0.05, P<0.01), and the scores of the primary symptoms and secondary symptoms of TCM, as well as the total score were lower than those of the control group (P<0.001, P<0.01). After treatment, the bilateral peak systolic velocity (PSV) and end diastolic velocity (EDV) were increased in the experimental group (P<0.001), the right EDV was increased in the control group (P<0.001), and the bilateral PSV and EDV in the experimental group were higher than those of the control group (P<0.05, P<0.01, P<0.001). After treatment, there was no significant difference in the total area of carotid plaque between the two groups. After treatment, there were no significant abnormalities in blood routine and liver and kidney functions in both groups, and the safety was good.

On the basis of conventional medication, the herbal cake-insulated moxibustion as an auxiliary treatment, presents its synergistic effect on CAS of interaction of phlegm and blood stasis syndrome in the patients, and this therapy is safe in clinical application.