Inhibition of autophagy suppresses osteogenic differentiation of stem cells from apical papilla.
10.12122/j.issn.1673-4254.2019.01.17
- Author:
Ying HUANG
1
;
Huacui XIONG
2
;
Ke CHEN
2
;
Xiaobin ZHU
2
;
Xiaoping YIN
3
;
Yun LIANG
2
;
Wei LUO
4
;
Qiyin LEI
4
Author Information
1. Stomatology Center, Shunde Hospital, Southern Medical University (The First People's Hospital of Shunde), Foshan 528308, China.
2. Stomatological Hospital, Southern Medical University, Guangzhou 510000, China.
3. Affiliated Hospital of Guilin Medical University, Guilin 541000, China.
4. Guangzhou Women and Children's Medical Center, Guangzhou 510623, China.
- Publication Type:Journal Article
- Keywords:
autophagy;
differentiation;
stem cells from apical papilla;
tumor necrosis factor-α
- MeSH:
Autophagy;
drug effects;
physiology;
Cell Differentiation;
drug effects;
physiology;
Cell Survival;
drug effects;
Cells, Cultured;
Dental Papilla;
cytology;
Green Fluorescent Proteins;
Humans;
Osteogenesis;
physiology;
Stem Cells;
drug effects;
physiology;
Transfection;
Tumor Necrosis Factor-alpha;
administration & dosage;
antagonists & inhibitors;
pharmacology
- From:
Journal of Southern Medical University
2019;39(1):106-112
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effects of autophagy on osteogenic differentiation of stem cells from the apical papilla (SCAPs) in the presence of tumor necrosis factor- (TNF-) stimulation .
METHODS:SCAPs treated with TNF- (0, 5, and 10 ng/mL) with or without 5 mmol/L 3-MA were examined for the expression of autophagy marker LC3-Ⅱ using Western blotting. The cells were transfected with GFP-LC3 plasmid and fluorescence microscopy was used for quantitative analysis of intracellular GFP-LC3; AO staining was used to detect the acidic vesicles in the cells. The cell viability was assessed with CCK-8 assays and the cell apoptosis rate was analyzed using flow cytometry. The cells treated with TNF- or with TNF- and 3-MA were cultured in osteogenic differentiation medium for 3 to 14 days, and real- time PCR was used to detect the mRNA expressions of osteogenesis-related genes (ALP, BSP, and OCN) for evaluating the cell differentiation.
RESULTS:TNF- induced activation of autophagy in cultured SCAPs. Pharmacological inhibition of TNF--induced autophagy by 3-MA significantly decreased the cell viability and increased the apoptosis rate of SCAPs ( < 0.05). Compared with the cells treated with TNF- alone, the cells treated with both TNF- and 3-MA exhibited decreased expressions of the ALP and BSP mRNA on days 3, 7 and 14 during osteogenic induction ( < 0.05) and decreased expression of OCN mRNA on days 3 and 7 during the induction ( < 0.05).
CONCLUSIONS:Autophagy may play an important role during the osteogenic differentiation of SCAPs in the presence of TNF- stimulation.
