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SIRT1/Atg5/autophagy are involved in the antiatherosclerosis effects of ursolic acid

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Abstract

The purpose of this study was to investigate the antiatherosclerosis effects of ursolic acid (UA) in high-fat diet-fed quails (Coturnix coturnix) and potential mechanism. Quails were treated with high-fat diet (14 % pork oil, 1 % cholesterol w/w) with or without UA (50, 150, or 300 mg/kg/day) for 10 weeks. Serum lipid profile was assessed at 0, 4.5, and 10 weeks. After 10 weeks, serum antioxidant status and morphology of aorta were assessed. Additionally, human umbilical vein endothelial cells (HUVECs) were exposed to 100 μg/ml oxidized low-density lipoprotein (ox-LDL) for 24 h, with or without pretreatment with UA (5, 10 or 20 μM) for 16 h, autophagy inhibitor 3-MA 5 mM for 2 h, or SIRT1 inhibitor EX-527 10 μM for 2 h. Cell viability and oxidative stress status were assessed and autophagy status was determined. Acetylation of lysine residue on Atg5 was assessed with immunoprecipitation. In results, high-fat diet negatively affected serum lipid profile and antioxidant status in quails and induced significant histological changes. Cotreatment with UA remarkably alleviated such changes. In HUVECs, ox-LDL treatment induced significant cytotoxicity along with oxidative stress, while UA cotreatment alleviated such changes significantly. UA treatment induced autophagy, enhanced SIRT1 expression, and decreased acetylation of lysine residue on Atg5. Cotreatment with 3-MA or EX-527 effectively abolished UA’s protective effects. In summary, UA exerted antiatherosclerosis effects in quails and protected HUVECs from ox-LDL induced cytotoxicity, and the mechanism is associated with increased SIRT1 expression, decreased Atg5 acetylation on lysine residue, and increased autophagy.

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Acknowledgments

The authors thank the affiliated hospital of Qingdao University for providing necessary instruments. This study was funded by the National Natural Science Foundation of China (Grant No. 81173593), received by Chunbo Wang.

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    Authors and Affiliations

    1. Department of Pharmacology, Qingdao University Medical College, Boya Building Room 422, 308 Ningxia Road, Qingdao, 266071, Shandong, China

      Qixiao Jiang, Ranran Hao, Hui Gao & Chunbo Wang

    2. The Institute of Human Nutrition Medical College, 38 Dengzhou Road, Qingdao, 266071, Shandong, China

      Wencheng Wang

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    1. Qixiao Jiang
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    Correspondence to Chunbo Wang.

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    Qixiao Jiang and Ranran Hao contributed equally to this work.

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    11010_2016_2787_MOESM1_ESM.tif

    Quantification method for the aorta histology. The areas of endothelium and/or smooth muscle that are rough and/or bulge into the aorta lumen were marked with black lines and indicated with arrows. ImageJ was used to measure the marked area. Atherosclerotic changes were quantified by normalizing the area with the total aorta area. Supplementary material 1 (TIFF 3040 kb)

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    Jiang, Q., Hao, R., Wang, W. et al. SIRT1/Atg5/autophagy are involved in the antiatherosclerosis effects of ursolic acid. Mol Cell Biochem 420, 171–184 (2016). https://doi.org/10.1007/s11010-016-2787-x

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