Erythropoietin Activates Autophagy to Regulate Apoptosis and Angiogenesis of Periodontal Ligament Stem Cells via the Akt/ERK1/2/BAD Signaling Pathway under Inflammatory Microenvironment

Author:

Huang Denghao12,Lei Jie12,Li Xingrui12,Jiang Zhonghao1,Luo Maoxuan1,Xiao Yao134ORCID

Affiliation:

1. Luzhou Key Laboratory of Orofacial Reconstruction and Regeneration, The Affiliated Stomatology Hospital of Southwest Medical University & Institute of Stomatology, Southwest Medical University, Luzhou, China

2. Cell Biology Technology Platform, Public Experiment Technology Center of Southwest Medical University, Luzhou, China

3. Department of Chengbei Outpatient, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou, China

4. Department of Orthodontics, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou, China

Abstract

Background. Angiogenic tissue engineering is a vital problem waiting to be settled for periodontal regeneration. Erythropoietin, a multieffect cytokine, has been reported as a protective factor for cell fate. According to our previous study, erythropoietin has a significantly angiogenic effect on periodontal ligament stem cells. To further explore its potential effects and mechanism, we studied biological behaviors of periodontal ligament stem cells under inflammatory microenvironment induced by different concentrations (0, 10, 20, 50, and 100 ng/mL) of tumor necrosis factor-α (TNF-α) and examined how different concentrations (0, 5, 10, 20, and 50 IU/mL) of erythropoietin changed biological behaviors of periodontal ligament stem cells. Materials and Methods. Cell Counting Kit-8 was used for cell proliferation assay. Annexin V-PI-FITC was used for cell apoptosis through flow cytometry. Matrigel plug was adopted to measure the angiogenic capacity in vitro. RNA sequencing was used to detect the downstream signaling pathway. Quantitative real-time polymerase chain reaction was conducted to examine mRNA expression level. Western blot and immunofluorescence were applied to testify the protein expression level. Results. Periodontal ligament stem cells upregulated apoptosis and suppressed autophagy and angiogenesis under inflammatory microenvironment. Erythropoietin could activate autophagy to rescue apoptosis and angiogenesis levels of periodontal ligament stem cells through the Akt/Erk1/2/BAD signaling pathway under inflammatory microenvironment. Conclusions. Erythropoietin could protect periodontal ligament stem cells from inflammatory microenvironment, which provided a novel theory for periodontal regeneration.

Funder

Ministry of Education of the People's Republic of China

Publisher

Hindawi Limited

Subject

Cell Biology,Molecular Biology

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