Henan Key Laboratory of Immunology and Targeted Therapy, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, Henan, China; Department of Biological and Chemical Engineering, Changsha University, Changsha 410022, Hunan, China.
Henan Key Laboratory of Immunology and Targeted Therapy, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, Henan, China.
Biochim Biophys Acta Mol Basis Dis. 2023 Dec;1869(8):166820. doi: 10.1016/j.bbadis.2023.166820. Epub 2023 Aug 7.
ATP synthase inhibitory factor 1 (ATPIF1) is a mitochondrial protein that regulates the activity of FF-ATP synthase. Mice lacking ATPIF1 throughout their bodies (Atpif1) exhibit a reduction in the number of neutrophils. However, it remains unclear whether the inactivation of ATPIF1 impairs the antibacterial function of mice, this study aimed to evaluate it using a mouse peritonitis model. Mice were intraperitoneally injected with E. coli to induce peritonitis, and after 24 h, the colonies of E. coli were counted in agarose plates containing mice peritoneal lavage fluids (PLF) or extract from the liver. Neutrophils were analyzed for glucose metabolism in glycolysis following LPS stimulation. Reactive oxygen species (ROS) and lactic acid (LA) levels in neutrophils were measured using flow cytometry and Seahorse analysis, respectively. N-Acetylcysteine (NAC) and 2-Deoxy-d-glucose (2-DG) were employed to assess the role of ROS and LA in neutrophil bactericidal activity. RNA-seq analysis was conducted in neutrophils to investigate potential mechanisms. In ATPIF1 neutrophils, bactericidal activity was enhanced, accompanied by increased levels of ROS and LA compared to wildtype neutrophils. The augmented bactericidal activity of ATPIF1 neutrophils was reversed by pretreatment with NAC or 2-DG. RNA-seq analysis revealed downregulation of multiple genes involved in glutathione metabolism, pyruvate oxidation, and heme synthesis, along with increased expression of inflammatory and apoptotic genes. This study suggests that the inactivation of the Atpif1 gene enhances glucose metabolism in neutrophils, resulting in increased bactericidal activity mediated by elevated levels of ROS and LA. Inhibiting ATPIF1 may be a potential approach to enhance antibacterial immunity.
ATP 合酶抑制因子 1（ATPIF1）是一种调节 FF-ATP 合酶活性的线粒体蛋白。全身缺乏 ATPIF1 的小鼠（Atpif1）表现出中性粒细胞数量减少。然而，目前尚不清楚 ATPIF1 的失活是否会损害小鼠的抗菌功能，本研究旨在使用小鼠腹膜炎模型对此进行评估。小鼠经腹腔内注射大肠杆菌诱导腹膜炎，24 小时后，在含有小鼠腹腔灌洗液（PLF）或肝脏提取物的琼脂糖平板上计数大肠杆菌菌落。用脂多糖刺激后分析中性粒细胞糖酵解中的葡萄糖代谢。使用流式细胞术和 Seahorse 分析分别测量中性粒细胞中活性氧（ROS）和乳酸（LA）的水平。使用 N-乙酰半胱氨酸（NAC）和 2-脱氧-D-葡萄糖（2-DG）评估 ROS 和 LA 在中性粒细胞杀菌活性中的作用。对中性粒细胞进行 RNA-seq 分析以研究潜在机制。在 ATPIF1 中性粒细胞中，与野生型中性粒细胞相比，杀菌活性增强，ROS 和 LA 水平升高。用 NAC 或 2-DG 预处理可逆转 ATPIF1 中性粒细胞的增强杀菌活性。RNA-seq 分析显示，与谷胱甘肽代谢、丙酮酸氧化和血红素合成相关的多个基因下调，同时炎症和凋亡基因表达增加。本研究表明，Atpif1 基因失活增强了中性粒细胞中的葡萄糖代谢，导致 ROS 和 LA 水平升高介导的杀菌活性增强。抑制 ATPIF1 可能是增强抗菌免疫的一种潜在方法。
