The mechanism of bone marrow-derived mesenchymal stem cells excessive senescence in severe aplastic anemia mouse model
10.3760/cma.j.issn.0253-2727.2017.04.012
- VernacularTitle: 重型再生障碍性贫血模型小鼠骨髓间充质干细胞过度衰老的机制研究
- Author:
Yiqing OU
1
;
Haiyan LIU
;
Wei LU
;
Mengjing WEN
;
Hong LIU
Author Information
1. Department of Hematology, Affiliated Hospital of Nantong University, Nantong 226001, China
- Publication Type:Journal Article
- Keywords:
Anemia, aplastic;
Mesenchymal stem cells;
Senescence
- From:
Chinese Journal of Hematology
2017;38(4):325-329
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the mechanism of excessive senescence in bone marrow-derived mesenchymal stem cells (BM-MSC) of mouse model with severe aplastic anemia (SAA) .
Methods:40 BALB/c mice were randomly assigned to two groups of control (n=20) and AA (n=20) . SAA mouse model was induced by intraperitoneal injection with IFN-γ and intragastric infusion with busulfan. BM-MSC were isolated and cultured from bone marrow of SAA and healthy mice. The cell morphology was observed by inverted microscope and cell cytoskeleton was stained by Rhodamine-Phalloidin; The level of proliferation was analyzed by CCK-8 method, and cell cycle was tested by flow cytometry. Senescence-associated β-galactosidase (SA-β-gal) assay was used to detect senescent BM-MSC; The expression of mTOR protein was detected by Western blot method.
Results:BM-MSC from normal mice presented spindle-shaped, clear boundaries and stress fibers were arranged in parallel, neat. while BM-MSCs from SAA mice presented cell volume increases, tiled, ill-shaped and the stress fiber appeared to be disordered. The decreased activity of proliferation [more cells restricted in G0/G1 phase [ (77.461±1.567) % vs (46.045±2.055) %, t=-34.384, P<0.001], increased percentage of SA-β-gal positive cells [ (75±11) % vs (28±8) %, t=15.454, P<0.001] and notably enhanced expression of mTOR of BM-MSC from SAA mice were observed when compared with those from normal mice.
Conclusion:This study clarified senescent BM-MSCs from SAA model mice, which could be caused by the excessive activation of mTOR pathway.
