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NOD2 induces VCAM-1 and ET-1 gene expression via NF-κB in human umbilical vein endothelial cells with muramyl dipeptide stimulation

NOD2 induziert VCAM-1- und ET-1-Genexpression über NF-κB in menschlichen Nabelvenenendothelzellen mit Muramyldipeptid-Stimulation

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Abstract

Objectives

Endothelial dysfunction is involved in various aspects of vascular biology and different stages of cardiovascular diseases (CVDs). Nucleotide-binding oligomerization domain-containing protein (NOD) 2, a pivotal innate immune receptor for muramyl dipeptide (MDP), has been reported to be a central regulator in CVDs. Previously, we reported that NOD2 played a leading role in MDP-triggered oxidative stress in endothelial cells (ECs). However, whether NOD2 participates in the regulatory mechanism of vascular cell adhesion molecule‑1 (VCAM-1) and endothelin‑1 (ET-1) expression was not elucidated.

Methods

Human umbilical vein endothelial cells (HUVECs) were stimulated with MDP for 12 h. mRNA expression of VCAM‑1 and ET‑1 was detected using real time polymerase chain reaction (PCR). Scrambled control small interfering RNA (siRNA) and NOD2 siRNA were transfected into HUVECs using Lipofectamine 2000 reagent (Invitrogen, Waltham, MA, USA). Furthermore, pyrrolidine dithiocarbamate was adopted to investigate the effect of nuclear factor κB (NF-κB) on NOD2-mediated VCAM‑1 and ET‑1 gene expression in MDP-treated HUVECs.

Results

Data showed that MDP significantly increased VCAM‑1 and ET‑1 mRNA expression, which was dependent on NOD2. In addition, NF-κB inhibition suppressed NOD2-mediated gene expression of VCAM‑1 and ET‑1.

Conclusion

Collectively, we confirmed NOD2 aggravated VCAM‑1 and ET‑1 gene expression through NF-κB in HUVECs treated with MDP.

Zusammenfassung

Zielsetzungen

Eine endotheliale Dysfunktion ist an vielfältigen Aspekten der Gefäßbiologie und an unterschiedlichen Stadien von Herz-Kreislauf-Erkrankungen (CVDs) beteiligt. NOD2 („nucleotide-binding oligomerization domain-containing protein“), ein entscheidender angeborener Immunrezeptor für Muramyldipeptid (MDP), wurde als zentraler Regulator bei Herz-Kreislauf-Erkrankungen beschrieben. In früheren Veröffentlichungen haben wir dargelegt, dass NOD2 eine wesentliche Funktion beim MDP-getriggerten oxidativen Stress in Endothelzellen (ECs) hat. Ob NOD2 jedoch am Regulationsmechanismus der Expression von VCAM‑1 („vascular cell adhesion molecule-1“) und ET‑1 („endothelin-1“) beteiligt ist, ließ sich nicht eindeutig klären.

Methoden

Humane Nabelvenenendothelzellen (HUVECs) wurden 12 h lang mit MDP stimuliert. Wir konnten die mRNA-Expression von VCAM‑1 und ET‑1 mittels Real-time-PCR (Polymerasekettenreaktion) nachweisen. „Scrambled control“ siRNA („small interfering RNA“) und NOD2-siRNA wurden mit Lipofectamine 2000-Reagenz (Invitrogen, Waltham/MA, USA) in HUVECs transfiziert. Ferner wurde Pyrrolidindithiocarbamat eingesetzt, um die Wirkung des nukleären Faktors κB (NF-κB) auf die NOD2-vermittelte VCAM-1- und ET-1-Genexpression in MDP-vorbehandelten HUVECs zu untersuchen.

Ergebnisse

Die Befunde zeigten, dass MDP die von NOD2 abhängige VCAM-1- und ET-1-mRNA-Expression signifikant erhöhte. Darüber hinaus supprimerte die NF-κB-Inhibition die NOD2-vermittelte Genexpression von VCAM‑1 und ET‑1.

Schlussfolgerung

Insgesamt bestätigten wir die durch NOD2 beeinträchtigte VCAM-1- und ET-1-Genexpression mittels NF-κB in mit MDP vorbehandelten HUVECs.

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Acknowledgements

The experiments were performed at Biological Medical Research Center of Zhongshan Hospital, Fudan University.

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Authors and Affiliations

  1. Department of Pharmacy, Shandong Provincial Hospital, Shandong First Medical University, 250021, Jinan, Shandong, China

    Ling-jun Kong

  2. Department of Anesthesiology, Peking University People’s Hospital, 100044, Beijing, China

    Ya-nan Wang

  3. Department of Clinical Pharmacy, Zhongshan Hospital, Fudan University, 200032, Shanghai, China

    Zi Wang & Qian-zhou Lv

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  1. Ling-jun Kong
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  2. Ya-nan Wang
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Correspondence to Ling-jun Kong or Qian-zhou Lv.

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Conflict of interest

L.-j. Kong, Y.-n. Wang, Z. Wang and Q.-Z. Lv declare that they have no competing interests.

For this article no studies with human participants or animals were performed by any of the authors. All studies performed were in accordance with the ethical standards indicated in each case.

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Kong, Lj., Wang, Yn., Wang, Z. et al. NOD2 induces VCAM-1 and ET-1 gene expression via NF-κB in human umbilical vein endothelial cells with muramyl dipeptide stimulation. Herz 46 (Suppl 2), 265–271 (2021). https://doi.org/10.1007/s00059-020-04996-y

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