State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China.
Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.
Br J Pharmacol. 2021 Mar;178(5):1055-1072. doi: 10.1111/bph.15339. Epub 2021 Feb 1.
Macrophage infiltration into the lungs is a characteristic of pulmonary hypertension (PH). Glycolysis is the main metabolic pathway for macrophage activation. However, the effect of macrophage glycolysis on the development of PH remains unknown. We investigated the effect of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKBF3), a critical enzyme of macrophage glycolysis, on PH development.
Lung tissues from PH patients were examined by immunostaining with macrophage markers. PH was induced in Wistar rats with SU5416/hypoxia and in mice with hypoxia. Lungs and macrophages were isolated for analysis by RT-PCR, western blot, flow cytometry, and immunostaining.
Expression of glycolytic molecules was increased in circulating peripheral blood mononuclear cells (PBMCs) and lung macrophages of PH patients. These results were also found in lung macrophages of SU5416/hypoxia (Su/Hx)-induced PH rats and hypoxia-induced PH mice. PH was ameliorated in myeloid-specific Pfkfb3-deficient mice (Pfkfb3 ) or mice treated with the PFKFB3 inhibitor 3PO, compared with their controls. Alveolar macrophages of PH Pfkfb3 mice produced lower levels of growth factors and pro-inflammatory cytokines than those of control mice. Circulating myeloid cells and lung myeloid cells were much fewer in PH Pfkfb3 mice than controls. Mechanistically, overexpression of Hif1a or Hif2a in bone marrow-derived macrophages (BMDMs) cultured with bone marrow of Pfkfb3 mice restored the decreased expression of pro-inflammatory cytokines and growth factors.
Myeloid Pfkfb3 deficiency protects mice from PH, thereby suggesting that myeloid PFKFB3 is one of the important targets in the therapeutic effect of PFKFB3 inhibition in PH treatment.
巨噬细胞浸润肺部是肺动脉高压（PH）的特征之一。糖酵解是巨噬细胞激活的主要代谢途径。然而，巨噬细胞糖酵解对 PH 发展的影响尚不清楚。我们研究了关键的巨噬细胞糖酵解酶 6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 3（PFKBF3）对 PH 发展的影响。
通过免疫染色用巨噬细胞标志物检测 PH 患者的肺组织。用 SU5416/缺氧诱导 Wistar 大鼠和缺氧诱导小鼠发生 PH。通过 RT-PCR、western blot、流式细胞术和免疫染色分析肺和巨噬细胞。
PH 患者循环外周血单核细胞（PBMC）和肺巨噬细胞中糖酵解分子的表达增加。SU5416/缺氧（Su/Hx）诱导的 PH 大鼠和缺氧诱导的 PH 小鼠的肺巨噬细胞中也发现了这些结果。与对照相比，髓样特异性 Pfkfb3 缺陷小鼠（Pfkfb3 ）或用 PFKFB3 抑制剂 3PO 治疗的 PH Pfkfb3 小鼠的 PH 得到改善。PH Pfkfb3 小鼠的肺泡巨噬细胞产生的生长因子和促炎细胞因子水平低于对照小鼠。PH Pfkfb3 小鼠的循环髓样细胞和肺髓样细胞比对照小鼠少得多。机制上，在骨髓来源的巨噬细胞（BMDM）中培养用 Pfkfb3 小鼠的骨髓时过表达 Hif1a 或 Hif2a 可恢复促炎细胞因子和生长因子表达的降低。
髓样细胞 Pfkfb3 缺失可保护小鼠免受 PH 影响，这表明髓样 PFKFB3 是 PFKFB3 抑制在 PH 治疗中的治疗效果的重要靶点之一。
