Bone marrow-derived mesenchymal stem cells modulate autophagy in RAW264.7 macrophages via the phosphoinositide 3-kinase/protein kinase B/heme oxygenase-1 signaling pathway under oxygen-glucose deprivation/restoration conditions.
Chin Med J (Engl)
; 134(6): 699-707, 2021 02 17.
Article
em En
| MEDLINE
| ID: mdl-33605598
ABSTRACT
BACKGROUND:
Autophagy of alveolar macrophages is a crucial process in ischemia/reperfusion injury-induced acute lung injury (ALI). Bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent cells with the potential for repairing injured sites and regulating autophagy. This study was to investigate the influence of BM-MSCs on autophagy of macrophages in the oxygen-glucose deprivation/restoration (OGD/R) microenvironment and to explore the potential mechanism.METHODS:
We established a co-culture system of macrophages (RAW264.7) with BM-MSCs under OGD/R conditions in vitro. RAW264.7 cells were transfected with recombinant adenovirus (Ad-mCherry-GFP-LC3B) and autophagic status of RAW264.7 cells was observed under a fluorescence microscope. Autophagy-related proteins light chain 3 (LC3)-I, LC3-II, and p62 in RAW264.7 cells were detected by Western blotting. We used microarray expression analysis to identify the differently expressed genes between OGD/R treated macrophages and macrophages co-culture with BM-MSCs. We investigated the gene heme oxygenase-1 (HO-1), which is downstream of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway.RESULTS:
The ratio of LC3-II/LC3-I of OGD/R treated RAW264.7 cells was increased (1.27â±â0.20 vs. 0.44â±â0.08, t = 6.67, Pâ <â0.05), while the expression of p62 was decreased (0.77â±â0.04 vs. 0.95â±â0.10, tâ=â2.90, Pâ <â0.05), and PI3K (0.40â±â0.06 vs. 0.63â±â0.10, tâ=â3.42, Pâ <â0.05) and p-Akt/Akt ratio was also decreased (0.39â±â0.02 vs. 0.58â±â0.03, tâ=â9.13, Pâ <â0.05). BM-MSCs reduced the LC3-II/LC3-I ratio of OGD/R treated RAW264.7 cells (0.68â±â0.14 vs. 1.27â±â0.20, tâ=â4.12, Pâ <â0.05), up-regulated p62 expression (1.10â±â0.20 vs. 0.77â±â0.04, tâ=â2.80, Pâ <â0.05), and up-regulated PI3K (0.54â±â0.05 vs. 0.40â±â0.06, tâ=â3.11, Pâ <â0.05) and p-Akt/Akt ratios (0.52â±â0.05 vs. 0.39â±â0.02, tâ=â9.13, Pâ <â0.05). A whole-genome microarray assay screened the differentially expressed gene HO-1, which is downstream of the PI3K/Akt signaling pathway, and the alteration of HO-1 mRNA and protein expression was consistent with the data on PI3K/Akt pathway.CONCLUSIONS:
Our results suggest the existence of the PI3K/Akt/HO-1 signaling pathway in RAW264.7 cells under OGD/R circumstances in vitro, revealing the mechanism underlying BM-MSC-mediated regulation of autophagy and enriching the understanding of potential therapeutic targets for the treatment of ALI.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Células-Tronco Mesenquimais
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article