Qiu Yuxuan, Li Linna, Guo Xiaohui, Liu Jiangwen, Xu Liang, Li Yanbo
Department of Endocrinology and Metabolic Disease, Harbin Medical University, Harbin, Heilongjiang 150076, P.R. China.
Department of Laboratory Medicine, Harbin City First Hospital, Harbin, Heilongjiang 150010, P.R. China.
Exp Ther Med. 2022 Apr;23(4):310. doi: 10.3892/etm.2022.11239. Epub 2022 Mar 1.
Evidence suggests that macrophage pyroptosis promotes the progression of diabetic atherosclerosis. Spermine, a natural cellular metabolite, demonstrates a protective effect against cardiovascular diseases. However, whether spermine has a protective effect against macrophage pyroptosis caused by high glucose (HG) and oxidized low-density lipoprotein (ox-LDL) conditions remains to be elucidated. To investigate the protective effect of spermine and the related underlying mechanism, THP-1 macrophages were treated with HG/ox-LDL, spermine, or the specific nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor ML385. Cell viability was detected using CCK-8, cell membrane permeability was analyzed using lactate dehydrogenase (LDH) and Hoechst/propidium iodide staining and pyroptosis-related gene and protein expression levels were evaluated using polymerase chain reaction and western blot analysis. Spermine showed a potent preventive effect on THP-1 macrophage pyroptosis and oxidative stress induced by HG/ox-LDL. Cells treated with spermine showed increased cell viability, reduced reactive oxygen species (ROS) production, decreased LDH levels in the supernatant and reduced cell swelling. In addition, spermine significantly reduced NLR family pyrin domain containing 3, cleaved caspase-1, N-gasdermin D and IL-1β expression, as well as IL-1β levels in the supernatant. This demonstrated that the inhibition of pyroptosis and oxidative stress due to spermine was Nrf2 dependent. Furthermore, spermine enhanced Nrf2 nuclear translocation, thereby increasing heme oxygenase-1 and NADPH quinone oxidoreductase-1 expression, which subsequently reduced ROS production. In addition, the anti-pyroptotic and antioxidant effects of spermine were reversed by ML385 inhibition of Nrf2. It was concluded that spermine prevented macrophage pyroptosis and increased ROS overproduction by activating the Nrf2 pathway. The data suggested that spermine may be a potential novel drug for the treatment of diabetic atherosclerosis because it targets macrophage pyroptosis.
有证据表明巨噬细胞焦亡会促进糖尿病动脉粥样硬化的发展。精胺作为一种天然的细胞代谢产物，对心血管疾病具有保护作用。然而，精胺是否对高糖（HG）和氧化型低密度脂蛋白（ox-LDL）诱导的巨噬细胞焦亡具有保护作用仍有待阐明。为了研究精胺的保护作用及其相关潜在机制，将THP-1巨噬细胞分别用HG/ox-LDL、精胺或特异性核因子红细胞2相关因子2（Nrf2）抑制剂ML385进行处理。使用CCK-8检测细胞活力，使用乳酸脱氢酶（LDH）以及Hoechst/碘化丙啶染色分析细胞膜通透性，并使用聚合酶链反应和蛋白质印迹分析评估焦亡相关基因和蛋白质的表达水平。精胺对HG/ox-LDL诱导的THP-1巨噬细胞焦亡和氧化应激具有显著的预防作用。用精胺处理的细胞显示出细胞活力增加、活性氧（ROS）生成减少、上清液中LDH水平降低以及细胞肿胀减轻。此外，精胺显著降低了含NLR家族pyrin结构域蛋白3、裂解的半胱天冬酶-1、N-气单胞菌溶素D和白细胞介素-1β的表达，以及上清液中白细胞介素-1β的水平。这表明精胺对焦亡和氧化应激的抑制作用依赖于Nrf2。此外，精胺增强了Nrf2的核转位，从而增加了血红素加氧酶-1和NADPH醌氧化还原酶-1的表达，随后减少了ROS的生成。此外，ML385抑制Nrf2可逆转精胺的抗焦亡和抗氧化作用。研究得出结论，精胺通过激活Nrf2途径预防巨噬细胞焦亡并增加ROS的过量产生。数据表明，精胺可能是一种潜在的新型药物，可用于治疗糖尿病动脉粥样硬化，因为它靶向巨噬细胞焦亡。
