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28 August 2019 : Animal Research  

High-Mobility Group Box 1 (HMGB1) Induces Migration of Endothelial Progenitor Cell via Receptor for Advanced Glycation End-Products (RAGE)-Dependent PI3K/Akt/eNOS Signaling Pathway

Yulong Zhang1ABCEF, Bo You12BCE, Xinzhu Liu1BCF, Jin Chen1BDF, Yizhi Peng1ADEF*, Zhiqiang Yuan1AEF

DOI: 10.12659/MSM.915829

Med Sci Monit 2019; 25:6462-6473

Abstract

BACKGROUND: High-mobility group box1 (HMGB1) is a cytokine that has been demonstrated to have an important role in inducing migration and homing of endothelial progenitor cells (EPCs) in the process of neovascularization during wound healing, but its specific mechanism remains elusive. The aim of this study was to investigate the effects of the HMGB-RAGE axis in EPC migration, as well as the underlying molecular mechanism responsible for these effects.

MATERIAL AND METHODS: EPCs were isolated from the mice and identified using flow cytometry and fluorescence staining. The effect of HMGB1 on the activity of EPCs was detected using the Cell Counting Kit-8 (CCK-8). Then, the migration of EPCs was detected by scratch wound-healing and cell migration assay. NO levels were analyzed by ELISA. The expression of p-PI3K, p-Akt, and p-eNOS was determined by Western blot analysis. RAGE expression was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot analysis. F-actin was assessed by fluorescent staining.

RESULTS: The results showed that HMGB1 induced a concentration-dependent migration of EPCs, and the migration was RAGE-dependent. The migration could be almost completely blocked by PI3K inhibitors and eNOS inhibitor. HMGB1-RAGE upregulated the expression of p-Akt, p-eNOS, and p-ERK. We also demonstrated that the MEK/ERK signaling pathway is not involved in the EPC migration induced by HMGB1-RAGE.

CONCLUSIONS: These data demonstrate that HMGB1 activates RAGE and induces PI3K/Akt/eNOS signaling transduction pathway activation to promote EPC migration. Therefore, the HMGB1-RAGE axis plays an important role in the EPC migration process and may become a potential target in wound healing.

Keywords: endothelial cells, HMGB1 Protein, Phosphatidylinositol 3-Kinases, Rage, Bone Marrow Cells, endothelial progenitor cells, Nitric Oxide Synthase Type III, Proto-Oncogene Proteins c-akt, Receptor for Advanced Glycation End Products

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750