Li Huan-Huan, Liu Xian, Wang Yu-Ping, Xu Xi, Zhu Lin, Zhang Wei, Ren Kun
College of Traditional Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230012, China.
College of Nursing, Anhui University of Chinese Medicine, Hefei 230012, China.
Metabolites. 2025 May 15;15(5):329. doi: 10.3390/metabo15050329.
Studies have shown that Atractylenolide I (AT-I) can exert anti-inflammatory and anti-oxidative effects, protecting against the development of various kinds of cardiovascular diseases. However, whether AT-I prevents nicotine-induced atherogenesis is unknown. This study was designed to explore the effects of AT-I on nicotine-induced macrophage pyroptosis and the progression of atherosclerosis. RT-qPCR and Western blot were used to detect the mRNA and protein levels of TXNIP and pyroptosis-related factors in THP-1-derived macrophages. ELISA was used to detect the secretion of pro-inflammatory cytokines. Hoechst/PI double-staining assay was used to assess plasma membrane integrity. The ROS assay kit, LDH release assay kit, and caspase-1 activity assay kit were used to detect ROS production, LDH release, and caspase-1 activity. Oil Red O, HE, and Masson staining were used to evaluate lipid accumulation, lesion size, and plaque stability in HFD-fed apoE mice. AT-I treatment significantly decreased pyroptosis-related factors expression, disrupted plasma membrane integrity, and down-regulated pro-inflammatory cytokines secretion, thereby inhibiting nicotine-induced pyroptosis of THP-1-derived macrophages. In addition, AT-I decreased ROS production and the expression of TLR4 and TXNIP. Lentivirus overexpression of TLR4 or TXNIP, or pre-treatment with ROS agonist, mainly reversed the anti-pyroptotic effects of AT-I in nicotine-treated THP-1-derived macrophages. Additionally, administering AT-I to HFD-fed apoE mice markedly decreased nicotine-induced up-regulation of pyroptosis-related proteins in the aortas. Enzymatic methods and ELISA assay suggested that AT-I improved dyslipidemia and inflammation in vivo. Oil Red O, HE, and Masson staining showed that AT-I alleviated lipid accumulation, decreased plaque size, and increased plaque stability. Taken together, AT-I can be regarded as a potential phytomedicine that protects against macrophage pyroptosis and atherosclerosis triggered by nicotine.
研究表明，白术内酯 I（AT-I）可发挥抗炎和抗氧化作用，预防各种心血管疾病的发生。然而，AT-I 是否能预防尼古丁诱导的动脉粥样硬化尚不清楚。本研究旨在探讨 AT-I 对尼古丁诱导的巨噬细胞焦亡及动脉粥样硬化进展的影响。采用 RT-qPCR 和蛋白质免疫印迹法检测 THP-1 来源巨噬细胞中 TXNIP 及焦亡相关因子的 mRNA 和蛋白质水平。采用酶联免疫吸附测定法检测促炎细胞因子的分泌。采用 Hoechst/PI 双染法评估质膜完整性。采用活性氧（ROS）检测试剂盒、乳酸脱氢酶（LDH）释放检测试剂盒和半胱天冬酶-1 活性检测试剂盒检测 ROS 生成、LDH 释放和半胱天冬酶-1 活性。采用油红 O、苏木精-伊红（HE）和 Masson 染色法评估高脂饮食喂养的载脂蛋白 E（apoE）小鼠的脂质蓄积、病变大小和斑块稳定性。AT-I 处理显著降低焦亡相关因子表达，破坏质膜完整性，下调促炎细胞因子分泌，从而抑制尼古丁诱导的 THP-1 来源巨噬细胞焦亡。此外，AT-I 降低了 ROS 生成以及 Toll 样受体 4（TLR4）和 TXNIP 的表达。TLR4 或 TXNIP 的慢病毒过表达，或用 ROS 激动剂预处理，主要逆转了 AT-I 对尼古丁处理的 THP-1 来源巨噬细胞的抗焦亡作用。此外，给高脂饮食喂养的 apoE 小鼠施用 AT-I 可显著降低尼古丁诱导的主动脉中焦亡相关蛋白的上调。酶法和酶联免疫吸附测定表明，AT-I 在体内改善了血脂异常和炎症。油红 O、HE 和 Masson 染色显示，AT-I 减轻了脂质蓄积，减小了斑块大小，并增加了斑块稳定性。综上所述，AT-I 可被视为一种潜在的植物药，可预防尼古丁引发的巨噬细胞焦亡和动脉粥样硬化。
