Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Shanghai Jiaotong University, Shanghai, China.
J Transl Med. 2021 Feb 10;19(1):62. doi: 10.1186/s12967-021-02727-3.
Atherosclerosis is a chronic vascular disease posing a great threat to public health. We investigated whether rosuvastatin (RVS) enhanced autophagic activities to inhibit lipid accumulation and polarization conversion of macrophages and then attenuate atherosclerotic lesions.
All male Apolipoprotein E-deficient (ApoE) mice were fed high-fat diet supplemented with RVS (10 mg/kg/day) or the same volume of normal saline gavage for 20 weeks. The burden of plaques in mice were determined by histopathological staining. Biochemical kits were used to examine the levels of lipid profiles and inflammatory cytokines. The potential mechanisms by which RVS mediated atherosclerosis were explored by western blot, real-time PCR assay, and immunofluorescence staining in mice and RAW264.7 macrophages.
Our data showed that RVS treatment reduced plaque areas in the aorta inner surface and the aortic sinus of ApoE mice with high-fat diet. RVS markedly improved lipid profiles and reduced contents of inflammatory cytokines in the circulation. Then, results of Western blot showed that RVS increased the ratio LC3II/I and level of Beclin 1 and decreased the expression of p62 in aortic tissues, which might be attributed to suppression of PI3K/Akt/mTOR pathway, hinting that autophagy cascades were activated by RVS. Moreover, RVS raised the contents of ABCA1, ABCG1, Arg-1, CD206 and reduced iNOS expression of arterial wall, indicating that RVS promoted cholesterol efflux and M2 macrophage polarization. Similarly, we observed that RVS decreased lipids contents and inflammatory factors expressions in RAW264.7 cells stimulated by ox-LDL, accompanied by levels elevation of ABCA1, ABCG1, Arg-1, CD206 and content reduction of iNOS. These anti-atherosclerotic effects of RVS were abolished by 3-methyladenine intervention. Moreover, RVS could reverse the impaired autophagy flux in macrophages insulted by chloroquine. We further found that PI3K inhibitor LY294002 enhanced and agonist 740 Y-P weakened the autophagy-promoting roles of RVS, respectively.
Our study indicated that RVS exhibits atheroprotective effects involving regulation lipid accumulation and polarization conversion by improving autophagy initiation and development via suppressing PI3K/Akt/mTOR axis and enhancing autophagic flux in macrophages.
动脉粥样硬化是一种慢性血管疾病，对公众健康构成极大威胁。我们研究了瑞舒伐他汀（RVS）是否通过增强自噬活性来抑制巨噬细胞中的脂质积累和极化转换，从而减轻动脉粥样硬化病变。
所有雄性载脂蛋白 E 缺陷（ApoE）小鼠均喂食高脂肪饮食，并补充 RVS（10mg/kg/天）或相同体积的生理盐水灌胃 20 周。通过组织病理学染色确定小鼠斑块的负担。使用生化试剂盒检测血脂谱和炎症细胞因子水平。通过 Western blot、实时 PCR 测定和免疫荧光染色在小鼠和 RAW264.7 巨噬细胞中探索 RVS 介导动脉粥样硬化的潜在机制。
我们的数据显示，RVS 治疗可减少高脂肪饮食喂养的 ApoE 小鼠主动脉内表面和主动脉窦的斑块面积。RVS 显著改善血脂谱并降低循环中炎症细胞因子的含量。然后，Western blot 结果显示，RVS 增加了 LC3II/I 比值和 Beclin 1 水平，并降低了主动脉组织中 p62 的表达，这可能归因于抑制了 PI3K/Akt/mTOR 通路，提示自噬级联反应被 RVS 激活。此外，RVS 提高了动脉壁 ABCA1、ABCG1、Arg-1、CD206 的含量，并降低了 iNOS 的表达，表明 RVS 促进了胆固醇流出和 M2 巨噬细胞极化。同样，我们观察到 RVS 降低了 ox-LDL 刺激的 RAW264.7 细胞中的脂质含量和炎症因子表达，同时 ABCA1、ABCG1、Arg-1、CD206 的水平升高，iNOS 的含量降低。这些 RVS 的抗动脉粥样硬化作用被 3-甲基腺嘌呤干预所消除。此外，RVS 可以逆转氯喹损伤的巨噬细胞中受损的自噬流。我们进一步发现，PI3K 抑制剂 LY294002 增强了 RVS 的自噬促进作用，激动剂 740 Y-P 则减弱了这种作用。
我们的研究表明，RVS 通过抑制 PI3K/Akt/mTOR 轴，改善自噬起始和发展，增强巨噬细胞中的自噬流，从而发挥抗动脉粥样硬化作用，涉及通过调节脂质积累和极化转换。
