AUTHOR=Zhang Yibo , Wang Xuelei , Gao Zhenyu , Li XuJie , Meng Ran , Wu Xiongfei , Ding Jie , Shen Weiliang , Zhu Junquan 

TITLE=Hypoxia-inducible factor-1α promotes macrophage functional activities in protecting hypoxia-tolerant large yellow croaker (Larimichthys crocea) against Aeromonas hydrophila infection

JOURNAL=Frontiers in Immunology

VOLUME=15

YEAR=2024

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1410082

DOI=10.3389/fimmu.2024.1410082

ISSN=1664-3224

ABSTRACT=<p>The immune system requires a high energy expenditure to resist pathogen invasion. Macrophages undergo metabolic reprogramming to meet these energy requirements and immunologic activity and polarize to M1-type macrophages. Understanding the metabolic pathway switching in large yellow croaker (<italic>Larimichthys crocea</italic>) macrophages in response to lipopolysaccharide (LPS) stimulation and whether this switching affects immunity is helpful in explaining the stronger immunity of hypoxia-tolerant <italic>L. crocea</italic>. In this study, transcript levels of glycolytic pathway genes (<italic>Glut1</italic> and <italic>Pdk1</italic>), mRNA levels or enzyme activities of glycolytic enzymes [hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), and lactate dehydrogenase A (LDHA)], aerobic respiratory enzymes [pyruvate dehydrogenase (PDH), isocitrate dehydrogenase (IDH), and succinate dehydrogenase (SDH)], metabolites [lactic acid (LA) and adenosine triphosphate (ATP)], levels of bactericidal products [reactive oxygen species (ROS) and nitric oxide (NO)], and transcripts and level changes of inflammatory factors [IL1β, TNFα, and interferon (IFN) γ] were detected in LPS-stimulated <italic>L. crocea</italic> head kidney macrophages. We showed that glycolysis was significantly induced, the tricarboxylic acid (TCA) cycle was inhibited, and metabolic reprogramming occurred, showing the Warburg effect when immune cells were activated. To determine the potential regulatory mechanism behind these changes, <italic>Lc</italic>HIF-1α was detected and found to be significantly induced and transferred to the nucleus after LPS stimulation. <italic>LcHif-1α</italic> interference led to a significant reduction in glycolytic pathway gene transcript expression, enzyme activity, metabolites, bactericidal substances, and inflammatory factor levels; a significant increase in the aerobic respiration enzymes; and decreased migration, invasion, and phagocytosis. Further ultrastructural observation by electron microscopy showed that fewer microspheres contained phagocytes and that more cells were damaged after <italic>LcHif-1α</italic> interference. <italic>LcHif-1α</italic> overexpression <italic>L. crocea</italic> head kidney macrophages showed the opposite trend, and promoter activities of <italic>Ldha</italic> and <italic>Il1β</italic> were significantly enhanced after <italic>LcHif-1α</italic> overexpression in HEK293T cells. Our data showed that <italic>Lc</italic>HIF-1α acted as a metabolic switch in <italic>L. crocea</italic> macrophages and was important in polarization. Hypoxia-tolerant <italic>L. crocea</italic> head kidney showed a stronger Warburg effect and inhibited the TCA cycle, higher metabolites, and bactericidal substance levels. These results collectively revealed that <italic>LcHif-1α</italic> may promote the functional activities of head kidney macrophages in protecting hypoxia-tolerant <italic>L. crocea</italic> from <italic>Aeromonas hydrophila</italic> infection.</p>