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Updated: Jun 4, 2026
Isolation of Primary Murine Brain Microvascular Endothelial Cells
Published on: November 14, 2014
NMN protects vascular endothelial cells from M1 macrophage-derived IL-1β-induced hyperpermeability by inhibiting
Takeshi Katayoshi1,2, Takahisa Nakajo1, Natsuko Kitajima1
1DHC Corporation Laboratories, Division 2, Chiba, Japan.
Background And Aims:
Vascular endothelial (VE) dysfunction, particularly endothelial hyperpermeability, is a critical pathological process in various inflammatory vascular diseases, including atherosclerosis, vasculitis, and sepsis. Nicotinamide mononucleotide (NMN), an NAD+ precursor, has shown anti-inflammatory and vascular protective effects in preclinical models. However, the mechanisms by which NMN preserves endothelial barrier integrity against macrophage-derived inflammatory stimuli remain unclear. This study examined the potential protective role of NMN in endothelial hyperpermeability induced by pro-inflammatory macrophages.
Methods:
A three-dimensional co-culture model of human umbilical vein endothelial cells (HUVECs) and M1 macrophages was constructed to reproduce inflammatory vascular microenvironments. Endothelial permeability was evaluated by measuring fluorescently labelled dextran and LDL passages from the luminal (top) to the abluminal side (bottom) of the insert.
Results:
M1 macrophage co-culture increased HUVEC permeability, and NMN pretreatment attenuated this hyperpermeability. Mechanistic analysis revealed that interleukin-1β (IL-1β) released by M1 macrophages was the primary contributor to endothelial hyperpermeability. NMN suppressed IL-1β-induced cell-cell gap formation and VE-cadherin degradation in HUVECs by inhibiting nuclear factor kappa B (NF-κB) pathway activation. These findings indicate that NMN prevents IL-1β-induced NF-κB activation and subsequent VE-cadherin degradation, thereby protecting against endothelial hyperpermeability caused by intercellular gap formation. Other NAD+ precursors, including nicotinamide riboside, similarly protected against IL-1β-induced hyperpermeability and VE-cadherin degradation. Alternatively, the NAD+-dependent deacetylase sirtuin 1 (SIRT1) inhibitor EX527 or SIRT1 siRNA knockdown abrogated NMN-mediated suppression of hyperpermeability and VE-cadherin degradation. Therefore, the protective effect of NMN against IL-1β-mediated endothelial dysfunction is dependent on the NAD+ -SIRT1 axis.
Conclusions:
This in vitro mechanistic study suggests that the NAD⁺-SIRT1 axis contributes to IL-1β-induced endothelial barrier disruption, supporting further investigation of NMN in inflammatory vascular diseases.
