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| Study on the mechanism of Bushen Jiangzhi Formula regulating autophagy and cholesterol efflux on the intervention of atherosclerosis in ApoE-/- mice#br# |
| YAN Li SHEN Dingzhu |
| Shanghai Geriatrics Institute of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200031, China |
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Abstract Objective To explore the mechanism of Bushen Jiangzhi Formula in regulating autophagy and cholesterol efflux in the intervention of atherosclerosis (AS). Methods One hundred ApoE-/- mice induced by high fat diet were divided into model group, kidney tonifying group, statin group and 3-methyladenine (3-MA) group by using random number table method, with 25 mice in each group. Twenty-five C57BL/6J mice were normal controls. Hematoxylin-eosin (HE), oil red O and FILIPIN staining were used to detect the pathological morphology, lipid accumulation and free cholesterol (FC) staining of the aortic sinus. Autophagosomes and lipid droplets were observed by electron microscopy. The content of serum total cholesterol (TC) and FC were detected by colorimetry. The expression of liver X receptor alpha (LXRα), ATP-binding membrane cassette transport protein A1 (ABCA1), microtubule-associated protein 1 light chain 3 (LC3) and p62 protein were detected by Western blot. Results The model group had more aortic sinus AS plaque, lipid accumulation and FC positive staining area than the normal group, and the differences were highly statistically significant (P ＜ 0.01). The normal group had intact aortic endothelial cell structure and autophagosome. The endothelial cells of aorta in model group were swollen with lipid droplets, but no autophagosomes were observed. The contents of TC, FC, and cholesterol ester (CE) in model group were higher than those in normal group, and the differences were highly statistically significant (P ＜ 0.01). The expression of LXRα, ABCA1, and LC3 Ⅱ/Ⅰ in aorta of model group was lower than that of normal group, and the expression of p62 was higher than that of normal group, the differences were highly statistically significant (P ＜ 0.01). AS plaque, lipid accumulation, and FC positive staining area of aortic sinus in the kidney tonifying group were less than those in the model group, and the differences were highly statistically significant (P ＜ 0.01). In the kidney tonifying group, autophagosomes were found in the endothelial cells of aorta. The contents of TC, FC, and CE in kidney tonifying group were lower than those in model group, and the differences were statistically significant (P ＜ 0.05 or P ＜ 0.01). The expression of LXRα, ABCA1, LC3Ⅱ/Ⅰ in kidney tonifying group was higher than that in model group, while the expression of p62 was lower than that in model group, the differences were statistically significant (P ＜ 0.05 or P ＜ 0.01). Conclusion Bushen Jiangzhi Formula may effectively intervene in the treatment of AS by regulating autophagy and cholesterol efflux.
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