基于整合素/YAP/JNK通路探讨隔药饼灸联合阿托伐他汀治疗ApoE-/-动脉粥样硬化小鼠的作用机制OA

Objective 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.Methods 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.Results 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).Conclusion 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.