PFKFB3 Inhibits Fructose Metabolism in Pulmonary Microvascular Endothelial Cells
PFKFB3 Inhibits Fructose Metabolism in Pulmonary Microvascular Endothelial Cells
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PFKFB3 抑制肺微血管内皮细胞中的果糖代谢
DOI：
10.1165/rcmb.2022-0443oc
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发表时间：
2023
影响因子：
6.4
通讯作者：
Koloteva, Anna
中科院分区：
文献类型：
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作者：
Lee, Ji Young;Stevens, Reece P.;Pastukh, Viktoriya V.;Pastukh, Viktor M.;Kozhukhar, Natalya;Alexeyev, Mikhail F.;Reisz, Julie A.;Nerguizian, David;D’Alessandro, Angelo;Koloteva, Anna
肺微血管内皮细胞有助于肺气体交换界面的完整性，并且它们具有高度糖酵解性。尽管葡萄糖和果糖代表可用于糖酵解的离散底物，但肺微血管内皮细胞更喜欢葡萄糖而不是果糖，并且这种选择所涉及的机制尚不清楚。 6-磷酸果糖-2-激酶/果糖-2, 6-双磷酸酶 3 (PFKFB3) 是一种重要的糖酵解酶，可驱动糖酵解通量对抗负反馈，并将糖酵解和果糖分解途径联系起来。我们假设 PFKFB3 抑制肺微血管内皮细胞中的果糖代谢。我们发现PFKFB3敲除细胞在缺氧条件下富含果糖的培养基中比野生型细胞存活得更好。海马测定、乳酸和葡萄糖测量以及稳定同位素示踪表明 PFKFB3 抑制果糖己糖激酶介导的糖酵解和氧化磷酸化。微阵列分析显示，果糖上调 PFKFB3，而 PFKFB3 敲除细胞会增加果糖特异性 GLUT5（葡萄糖转运蛋白 5）的表达。使用条件性内皮特异性 PFKFB3 敲除小鼠，我们证明内皮 PFKFB3 敲除可增加果糖灌胃后肺组织乳酸的产生。最后，我们发现肺炎会增加 ICU 机械通气患者 BAL 液中的果糖含量。因此，PFKFB3敲除增加了肺微血管内皮细胞中GLUT5的表达和己糖激酶介导的果糖使用，从而促进其存活。我们的研究结果表明，PFKFB3 是一种分子开关，可以控制糖酵解中葡萄糖与果糖的使用，并有助于更好地了解呼吸衰竭期间的肺内皮细胞代谢。
Pulmonary microvascular endothelial cells contribute to the integrity of the lung gas exchange interface, and they are highly glycolytic. Although glucose and fructose represent discrete substrates available for glycolysis, pulmonary microvascular endothelial cells prefer glucose over fructose, and the mechanisms involved in this selection are unknown. 6-Phosphofructo-2-kinase/fructose-2, 6-bisphosphatase 3 (PFKFB3) is an important glycolytic enzyme that drives glycolytic flux against negative feedback and links glycolytic and fructolytic pathways. We hypothesized that PFKFB3 inhibits fructose metabolism in pulmonary microvascular endothelial cells. We found that PFKFB3 knockout cells survive better than wild-type cells in fructose-rich medium under hypoxia. Seahorse assays, lactate and glucose measurements, and stable isotope tracing showed that PFKFB3 inhibits fructose–hexokinase–mediated glycolysis and oxidative phosphorylation. Microarray analysis revealed that fructose upregulates PFKFB3, and PFKFB3 knockout cells increase fructose-specific GLUT5 (glucose transporter 5) expression. Using conditional endothelial-specific PFKFB3 knockout mice, we demonstrated that endothelial PFKFB3 knockout increases lung tissue lactate production after fructose gavage. Last, we showed that pneumonia increases fructose in BAL fluid in mechanically ventilated ICU patients. Thus, PFKFB3 knockout increases GLUT5 expression and the hexokinase-mediated fructose use in pulmonary microvascular endothelial cells that promotes their survival. Our findings indicate that PFKFB3 is a molecular switch that controls glucose versus fructose use in glycolysis and help better understand lung endothelial cell metabolism during respiratory failure.