Xiong Wangqiong, Fei Minzhong, Wu Chuntao, Wang Wei, Luo Rong, Shen Liping, Zhang Zheng
ECG Room, Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University, Shanghai 201700, P.R. China.
Cardiovascular Department, Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University, Shanghai 201700, P.R. China.
Exp Ther Med. 2020 Mar;19(3):2266-2272. doi: 10.3892/etm.2019.8379. Epub 2019 Dec 27.
Effect of atorvastatin inhibition of endoplasmic reticulum stress and amelioration of atherosclerosis through AMPK pathway were studied. Eight-week-old male apolipoprotein E-deficient (ApoE/) mice were fed with high-fat diet for 2 weeks and randomly divided into two groups: Atorvastatin treatment group was given atorvastatin (5 mg/kg/day) injection for a total of 6 weeks; control group was given the same dose of PBS through intraperitoneal injection for a total of 6 weeks. H&E staining was used to detect plaque size; immunohistochemical staining was used to detect T cells, macrophages and phospho-protein kinase-like ER kinase (phospho-PERK) in localized plaques. Proteins were extracted from mouse thoracic and abdominal aortic tissues. Western blot analysis was used to detect the protein expression levels of endoplasmic reticulum stress-related molecules phospho-eukaryotic initiation factor-2α (p-eIF2α), eukaryotic initiation factor (eIF2a), and sliced x-box binding protein 1 (sXBP-1). Cultured human umbilical vein endothelial cells (HUVECs), induced endoplasmic reticulum stress with human oxidized low density lipoprotein (ox-LDL), were treated with atorvastatin, AMPK agonist 5-amino-4-imidazolecarboxamide riboside-I-β-D-ribofuranoside (AICAR) and AMPK-DN that expressed a dominant-negative mutant of AMPK. Western blot analysis was used to test the expression levels of endoplasmic reticulum stress-related molecules p-elF2a and sXBP-1. The area of aortic plaques in atorvastatin group was obviously decreased, and the infiltrations of CD3 T cells and macrophages in the localized plaques were reduced. The endoplasmic reticulum stress-related proteins sXBP-1 and p-eIF2a were significantly reduced. The results of immunohistochemistry also showed a significant decrease in the level of phospho-PERK (p-PERK) in atorvastatin group. The results in ox-LDL-induced HUVECs showed that atorvastatin inhibited ox-LDL-induced endoplasmic reticulum stress, and the AMPK agonist AICAR also had the same effect, which was offset by DN-AMPK treatment. Atorvastatin inhibits ER stress both and and this protective effect is mediated by AMPK activation.
研究了阿托伐他汀通过AMPK途径抑制内质网应激及改善动脉粥样硬化的作用。将8周龄雄性载脂蛋白E缺陷（ApoE-/-）小鼠高脂喂养2周后随机分为两组：阿托伐他汀治疗组给予阿托伐他汀（5 mg/kg/天）注射，共6周；对照组给予相同剂量的PBS腹腔注射，共6周。采用苏木精-伊红（H&E）染色检测斑块大小；采用免疫组织化学染色检测局部斑块中的T细胞、巨噬细胞和磷酸化蛋白激酶样内质网激酶（phospho-PERK）。从小鼠胸主动脉和腹主动脉组织中提取蛋白质。采用蛋白质印迹分析检测内质网应激相关分子磷酸化真核起始因子-2α（p-eIF2α）、真核起始因子（eIF2α）和剪接型X盒结合蛋白1（sXBP-1）的蛋白表达水平。用人类氧化低密度脂蛋白（ox-LDL）诱导培养的人脐静脉内皮细胞（HUVECs）发生内质网应激，分别用阿托伐他汀、AMPK激动剂5-氨基-4-咪唑甲酰胺核糖苷-I-β-D-呋喃核糖苷（AICAR）和表达AMPK显性负性突变体的AMPK-DN进行处理。采用蛋白质印迹分析检测内质网应激相关分子p-elF2α和sXBP-1的表达水平。阿托伐他汀组主动脉斑块面积明显减小，局部斑块中CD3+T细胞和巨噬细胞浸润减少。内质网应激相关蛋白sXBP-1和p-eIF2α明显降低。免疫组织化学结果也显示阿托伐他汀组磷酸化PERK（p-PERK）水平显著降低。在ox-LDL诱导的HUVECs中的结果表明，阿托伐他汀抑制ox-LDL诱导的内质网应激，AMPK激动剂AICAR也有相同作用，而DN-AMPK处理可抵消这种作用。阿托伐他汀通过激活AMPK抑制内质网应激，且这种保护作用是由AMPK激活介导的。
