Mechanism of herbal cake-separated moxibustion combined with atorvastatin in treating ApoE^-/-atherosclerotic mice based on the integrin/YAP/JNK signaling pathway

ObjectiveTo 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.MethodsMale 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.ResultCompared 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）.ConclusionHerbal 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.