目的 研究茱萸丸对动脉粥样硬化（atherosclerosis，AS）的影响及其相关机制。方法 采用高脂饲料喂养雄性低密度脂蛋白受体敲除（low density lipoprotein receptor knockout，LDLR^-/-）小鼠诱导AS模型，造模周期为12周。造模成功的50只LDLR^-/-小鼠随机分成模型组及茱萸丸低、中、高剂量（130.54、261.08、552.16 mg/kg）组和阿托伐他汀（10.40 mg/kg）组，每组10只，C57BL/6J小鼠10只作为对照组。给药12周后，采用生化法检测小鼠血清三酰甘油（triglyceride，TG）、总胆固醇（total cholesterol，TC）、低密度脂蛋白胆固醇（low density lipoprotein cholesterol，LDL-C）、高密度脂蛋白胆固醇（high density lipoprotein cholesterol，HDL-C）水平；苏木素-伊红（hematoxylin-eosin，HE）、Masson、油红O染色观察小鼠主动脉的病理学改变；ELISA检测血清中白细胞介素-18（interleukin-18，IL-18）、IL-1β、活性氧（reactive oxygen species，ROS）水平和超氧化物歧化酶（superoxide dismutase，SOD）活性；超高液相色谱串联质谱法（ultra-high performance liquid chromatography-tandem mass spectrometry，UHPLC-MS/MS）检测小鼠血浆中三甲胺（trimethylamine，TMA）、氧化三甲胺（trimethylamine oxide，TMAO）含量；16S rRNA技术检测小鼠粪便中肠道菌群物种组成差异；荧光探针法检测主动脉ROS的荧光强度；免疫荧光及Western blotting检测小鼠主动脉硫氧还蛋白相互作用蛋白（thioredoxin interacting protein，TXNIP）和核苷酸结合寡聚化结构域样受体蛋白（NOD like receptor family pyrin domain containing 3，NLRP3）蛋白表达；qRT-PCR检测主动脉硫氧还蛋白（thioredoxin，TRX）、TXNIP、凋亡相关斑点样蛋白（apoptosis-associated speck-like protein，ASC）、NLRP3、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）mRNA表达。结果 与对照组比较，模型组小鼠血清TC、TG、LDL-C水平升高（P＜0.01），HDL-C水平显著下降（P＜0.01），主动脉内膜大面积增厚，形成明显的泡沫细胞，脉壁胶原蛋白沉积增多，血清中IL-18、IL-1β、ROS、TMA、TMAO水平显著升高（P＜0.01），SOD活性显著降低（P＜0.01），Chao1、Ace以及observed species指数降低（P＜0.01），拟杆菌门、粪杆菌属、布劳特氏属、乳杆菌属菌群丰度显著下降（P＜0.01），主动脉中ROS含量增加（P＜0.01），TXNIP、NLRP3蛋白与mRNA表达水平升高（P＜0.01），TRX、ASC、Caspase-1 mRNA表达水平升高（P＜0.01）。与模型组比较，茱萸丸组血清TG、TC、LDL-C水平显著降低（P＜0.05、0.01），HDL-C水平显著升高（P＜0.01），主动脉斑块、血管壁泡沫细胞及胶原蛋白沉积均呈现不同程度的改善，血清中IL-18、IL-1β、ROS、TMA、TMAO水平显著降低（P＜0.05、0.01），SOD活性显著升高（P＜0.01），Chao1、Ace以及observed species指数均明显升高（P＜0.01），拟杆菌门、粪杆菌属、布劳特氏属、乳杆菌属菌群丰度升高（P＜0.05、0.01），厚壁菌门菌群丰度降低（P＜0.05），主动脉中ROS含量减少（P＜0.05、0.01），TXNIP、NLRP3蛋白与mRNA表达水平显著降低（P＜0.01），TRX、ASC、Caspase-1 mRNA表达水平显著降低（P＜0.01）。结论 茱萸丸能够显著抑制LDLR^-/- AS小鼠主动脉病理改变、调节肠道菌群中有益菌和有害菌的相对丰度，减少TMAO生成，抑制ROS/TXNIP/NLRP3信号通路，改善炎症反应，减轻内皮细胞损伤，从而发挥抗AS的作用。

Objective To investigate the effect and mechanism of Zhuyu Pills (茱萸丸) on atherosclerosis (AS). Methods Male low-density lipoprotein receptor knockout (LDLR^−/−) mice were fed high-fat diet to induce AS model. The modeling period was 12 weeks. The 50 LDLR^−/− mice successfully modeled were randomly divided into model group, Zhuyu Pills low-, medium- and high-dose (130.54, 261.08, 552.16 mg/kg) group and atorvastatin (10.40 mg/kg) group, with 10 mice in each group. Another 10 C57BL/6J mice were set as control group. After 12 weeks of administration, triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) levels in serum of mice were detected by biochemical method; The pathological changes of aorta were observed by hematoxylin-eosin (HE) staining, Masson staining and oil red O staining; The contents of interleukin-18 (IL-18), IL-1β, reactive oxygen species (ROS) and superoxide dismutase (SOD) were detected by ELISA; The contents of trimethylamine (TMA) and trimethylamine oxide (TMAO) in serum of mice were detected by ultra-high liquid chromatography tandem mass spectrometry (UHPLC-MS/MS); The species composition of intestinal flora in stool of mice was detected by 16S rRNA technique; The fluorescence intensity of ROS in aorta was detected by fluorescence probe; The expressions of thioredoxin interacting protein (TXNIP) and NOD like receptor family pyrin domain containing 3 (NLRP3) in aorta was detected by immunofluorescence and Western blotting; The mRNA expressions of thioredoxin (TRX), TXNIP, apoptosis-associated speck-like protein (ASC), NLRP3 and cystein-asparate protease-3 (Caspase-1) were detected by qRT-PCR. Results Compared with control group, TC, TG and LDL-C levels in serum of mice in model group were significantly increased (P < 0.01), HDL-C level was decreased (P < 0.01), the aortic intima was thickened in a large area and formed obvious foam cells, the collagen deposition in the pulse wall was increased, IL-18, IL-1β, ROS, TMA and TMAO levels in serum were increased (P < 0.01), SOD activity was decreased (P < 0.01). The index of Chao1, Ace and observed species were decreased (P < 0.01), the abundance of Bacteroidota, Faecalibaculum, Blautia and Lactobacillus were significantly decreased (P < 0.01), ROS levels in aorta was increased (P < 0.01), and the expression levels of TXNIP and NLRP3 protein and mRNA expressions were increased, the mRNA expression levels of TRX, ASC and Caspase-1 were increased (P < 0.01). Compared with model group, TG, TC and LDL-C levels in serum were decreased (P < 0.05, 0.01), HDL-C level was increased in Zhuyu Pills group, aortic plaque, vascular wall foam cells and collagen deposition were improved to varying degrees, IL-18, IL-1β, ROS, TMA and TMAO levels in serum were decreased (P < 0.05, 0.01), and SOD activity was increased (P < 0.01). Chao1, Ace and observed species indexes were significantly increased (P < 0.01), the abundance of Bacteroidota, Faecalibaculum, Blautia and Lactobacillus were increased (P < 0.05, 0.01), while the abundance of Firmicutes was decreased (P < 0.05), ROS level in aorta was decreased (P < 0.05, 0.01), protein and mRNA expression levels of TXNIP and NLRP3 were decreased (P < 0.01), mRNA expression levels of TRX, ASC and Caspase-1 were decreased (P < 0.01). Conclusion Zhuyu Pills can significantly inhibit the pathological changes of aorta in LDLR^−/− AS mice, regulate the relative abundance of beneficial and harmful bacteria in intestinal flora, reduce TMAO production, inhibit ROS/TXNIP/NLRP3 signaling pathway, improve inflammatory response, and alleviate endothelial cell damage, thus playing an anti-AS role.

国家自然科学基金面上项目（81973743）；四川省科技厅科技创新创业人才和苗子工程项目（20MZGC0241）；四川省科技厅重点研发项目（2022YFS0381）