PKM2 regulates angiogenesis of VR‐EPCs through modulating glycolysis, mitochondrial fission, and fusion

Abstract

Vascular resident endothelial progenitor cells (VR-EPCs) have a certain ability to differentiate into endothelial cells (ECs) and participate in the process of angiogenesis. Glycolysis and mitochondrial fission and fusion play a pivotal role in angiogenesis. Pyruvate kinase muscle isoenzyme 2 (PKM2), which mediates energy metabolism and mitochondrial morphology, is regarded as the focus of VR-EPCs angiogenesis in our study. VR-EPCs were isolated from the hearts of 12-weeks-old Sprague-Dawley rats. The role of PKM2 on angiogenesis was evaluated by tube formation assay, wound healing assay, transwell assay, and chick chorioallantoic membrane assay. Western blot analysis, flow cytometry, mitochondrial membrane potential detection, reactive oxygen species (ROS) detection, immunofluorescence staining, and quantitative real-time polymerase chain reaction were used to investigate the potential mechanism of PKM2 for regulating VR-EPCs angiogenesis.We explored the function of PKM2 on the angiogenesis of VR-EPCs. DASA-58 (the activator of PKM2) promoted VR-EPCs proliferation and PKM2 activity, it also could promote angiogenic differentiation. At the same time, DASA-58 significantly enhanced glycolysis, mitochondrial fusion, slightly increased mitochondrial membrane potential, and maintained ROS at a low level. C3k, an inhibitor of PKM2, inhibited PKM2 activity, expression of angiogenesis-related genes and tube formation. Besides, C3k drastically reduced glycolysis and mitochondrial membrane potential while significantly promoting mitochondrial fission and ROS level.Activation of PKM2 could promote VR-EPCs angiogenesis through modulating glycolysis, mitochondrial fission and fusion. By contrast, PKM2 inhibitor has opposite effects.

Mitochondria Lead the Way: Mitochondrial Dynamics and Function in Cellular Movements in Development and Disease

“…Glycolysis in the EC tip cells is essential for their differentiation whereas OXPHOS in non-tip ECs plays a role in cell survival, transcriptional activities and ROS production ( Yetkin-Arik et al, 2019 ). Mitochondria in the ECs become elongated upon proangiogenic stimuli by the upregulation of Mfn ( Lugus et al, 2011 ; Zhou et al, 2015 ; Ren et al, 2020 ) and Opa1 activity ( Xu et al, 2019 ; Herkenne et al, 2020 ) and downregulation of Drp1 and Fis1 ( Xu et al, 2019 ; Ren et al, 2020 ). Opa1 also regulates the activity of NF-ƙB and is essential for the proliferation and migration of ECs in vitro ( Herkenne et al, 2020 ).…”

PERK controls bone homeostasis through the regulation of osteoclast differentiation and function

“…Western Blot was conducted as described previously 61 . After treated, BMMs were lysed by using RIPA Buffer (Boster) containing phosphatase inhibitor and protease inhibitor to extract total protein.…”

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