Xuefu Zhuyu Capsules Ameliorate Atherosclerosis in Mice by Regulating Sirt3/EPAC1 Signaling Pathway
10.13422/j.cnki.syfjx.20241622
- VernacularTitle:血府逐瘀胶囊通过调控Sirt3/EPAC1信号通路对动脉粥样硬化小鼠的影响
- Author:
Bo YAO
1
;
Hengwen CHEN
2
;
Jiyu GONG
1
;
Xuanhui HE
2
Author Information
1. School of Pharmaceutical Sciences， Changchun University of Chinese Medicine， Changchun 130117， China
2. Guang'anmen Hospital， China Academy of Chinese Medical Sciences， Beijing 100053， China
- Publication Type:Journal Article
- Keywords:
Xuefu Zhuyu capsules;
atherosclerosis;
silencing regulatory factor 2-like protein 3 （Sirt3）;
exchange protein directly activated by cAMP 1 （EPAC1）;
mitochondria
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(21):31-41
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo observe the effects of Xuefu Zhuyu capsules （XFZY） on blood lipid levels and aortic plaques in the mouse model of atherosclerosis （AS） induced by a high-fat diet by regulating the silencing regulatory factor 2-like protein 3 （Sirt3）/exchange protein directly activated by cAMP 1 （EPAC1） signaling pathway and explore the mechanism of XFZY in ameliorating AS. MethodMice were assigned into normal， model， blank， rosuvastatin （0.05 g·kg-1·d-1）， and low-， medium-， and high-dose （0.3， 0.6， 1.2 g·kg-1·d-1， respectively） XFZY groups. The normal group consisted of normal C57BL/6J mice， while the other groups consisted of ApoE-/- C57BL/6J mice. The normal group and blank group were fed routinely， and the rest groups were fed with a high-fat diet for 24 consecutive weeks for the modeling of AS. The drug intervention groups were administrated with corresponding drugs by gavage， and model group and blank group with an equal volume of deionized water for 6 consecutive weeks. The small animal B-ultrasound was used to evaluate the mouse heart function and aortic plaque condition. A fully automated biochemical analyzer was used to measure the levels of blood lipids such as total cholesterol （CHOL）， triglycerides （TG）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and extremely low-density lipoprotein （VLDL） in mice. Oil red O staining was employed to observe lipid deposition in the aorta. Hematoxylin-eosin staining and Masson staining were employed to observe the pathological changes and collagen deposition in mouse blood vessels. Transmission electron microscopy was employed to observe the mitochondrial damage in mouse aorta. The levels of adrenocorticotropic hormone （ACTH）， adenosine triphosphate （ATP）， and nicotinic choline receptor α1 （CHRNα1）， and total superoxide dismutase （T-SOD） were measured by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were performed to determine the mRNA and protein levels， respectively， of Sirt3， EPAC1， Caspase-3， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the mouse aorta and heart. ResultMultiple AS plaques were observed in the aortic arch， indicating that the model was successfully established. Compared with the model group， the XFZY groups showed reduced and narrowed plaques. Compared with the normal group， the model group showed elevated CHOL level （P<0.01）. Compared with the model group， rosuvastatin and low-dose XFZY lowered the CHOL and TG levels （P<0.01）. Compared with the normal group， the model group presented a large number of protruding red lipid plaques on the aortic wall and increased percentage of AS plaque area to total tissue area （P<0.01）. Compared with the model group， low-dose XFZY reduced the plaque load （P<0.05）. Compared with the model group， XFZY at different doses reduced the lipid plaques and collagen deposition. Compared with the normal group， the model group showed decreased or disappeared mitochondrial cristae and presented severe damage of the membrane structure in endothelial cells. The mitochondria of endothelial cells in each treatment group approached the normal structure， with mitochondrial cristae faintly visible. Compared with the normal group， the model group showcased reduced myocardial mitochondrial ATP activity （P<0.01）， which were rescored in the drug intervention groups （P<0.01）. Compared with the normal group， the modeling inhibited the expression of Sirt3 （P<0.01） and promoted the expression of EPAC1 （P<0.01）. Compared with the model group， low-dose XFZY increased the Sirt3 content （P<0.01） and medium-dose XFZY increased the EPAC1 content （P<0.01）， which indicated that XFZY treatment upregulated the mRNA and protein levels of Sirt3 and downregulated the mRNA and protein levels of EPAC1. ConclusionXFZY can alleviate the aortic lipid deposition， reduce the AS plaque area， improve the mitochondrial morphology and functions in endothelial cells， increase the ATP activity， upregulate the expression of Sirt3， and downregulate the expression of EPAC1 in AS mice by regulating mitochondrial energy metabolism via the Sirt3/EPAC1 signaling pathway.
