Hyperactivation of PI3K/AKT/mTOR signal pathway impairs TNF-α-induced autophagy in mesenchymal stem cells from patients with ankylosing spondylitis.
10.12122/j.issn.1673-4254.2022.02.15
- Author:
Zhen Hua LIU
1
;
Shao Xiong MIN
2
;
Xiu Yi LU
3
;
Shui Zhong CEN
1
;
Zhi Peng CHEN
4
;
Tao WANG
1
;
Jian Jun LI
1
;
Wei Bo ZENG
1
;
Su Jun QIU
1
Author Information
1. Department of Orthopedics, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
2. Department of Orthopedics, Shenzhen Hospital, Peking University, Shenzhen 518034, China.
3. Department of Dermatology, Fourth Affiliated Hospital of Guangzhou Medical University, Guangzhou 510316, China.
4. Department of Orthopedics, Sun Yat-sen Memorial Hospital, Sun Yat- sen University, Guangzhou 510120, China.
- Publication Type:Journal Article
- Keywords:
ankylosing spondylitis;
autophagy;
mesenchymal stem cells;
tumor necrosis factor-α
- MeSH:
Autophagy;
Humans;
Mesenchymal Stem Cells/metabolism*;
Phosphatidylinositol 3-Kinases/metabolism*;
Proto-Oncogene Proteins c-akt/metabolism*;
Signal Transduction;
Spondylitis, Ankylosing;
TOR Serine-Threonine Kinases/metabolism*;
Tumor Necrosis Factor-alpha/metabolism*
- From:
Journal of Southern Medical University
2022;42(2):272-277
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the changes in autophagy of mesenchymal stem cells (MSCs) from patients with ankylosing spondylitis and explore the mechanism for decreased autophagy in ASMSCs.
METHODS:MSCs collected from 14 patients with AS (ASMSCs) and from 15 healthy donors (HDMSCs) were cultured in the absence or presence of 25 ng/mL TNF-α for 6 h. Autophagy of the cells was determined by immunofluorescence staining of GFP-LC3B, and the results were confirmed by detecting the protein expressions of autophagy markers LC3 II/LC3 I and P62. The mRNA expressions of the related genes were detected using qRT-PCR, and the protein expressions of the autophagy markers and signaling pathway-related molecules were determined with Western blotting. TG100713 was used to block the PI3K/AKT/mTOR signal pathway, and its effect on autophagy of ASMSCs was evaluated.
RESULTS:ASMSCs showed significantly weaker GFP-LC3B puncta staining and lower protein expression levels of LC3 II/LC3 I but higher levels of P62 protein (P < 0.05), indicating a decreased autophagy capacity as compared with HDMSCs. TNF-α-induced ASMSCs showed significantly higher protein expressions of p-PI3K/ PI3K, p-AKT/AKT and p-mTOR/mTOR than HDMSCs (P < 0.05), suggesting hyperactivation of the PI3K/AKT/mTOR signaling pathway in ASMSCs. Blocking PI3K/AKT/mTOR signaling with TG100713 eliminated the difference in TNF-α-induced autophagy between HDMSCs and ASMSCs.
CONCLUSION:In patients with AS, hyperactivation of the PI3K/AKT/mTOR signaling pathway results in decreased autophagy of the MSCs and potentially contributes to chronic inflammation.
