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Mesenchymal Stromal Cells Accelerate Epithelial Tight Junction Assembly via the AMP-Activated Protein Kinase Pathway, Independently of Liver Kinase B1
- Source :
- Stem Cells International, Stem Cells International, 2017, Stem Cells International, Hindawi Publishing Corporation, 2017, Stem Cells International, Vol 2017 (2017)
- Publication Year :
- 2017
- Publisher :
- HAL CCSD, 2017.
-
Abstract
- Background. Mesenchymal stromal cells (MSC) are fibroblast-like multipotent cells capable of tissue-repair properties. Given the essentiality of tight junctions (TJ) in epithelial integrity, we hypothesized that MSC modulate TJ formation, via the AMP-activated kinase (AMPK) pathway. Liver kinase-β1 (LKB1) and Ca2+-calmodulin-dependent protein kinase kinase (CaMKK) represent the main kinases that activate AMPK.Methods. The in vitro Ca2+switch from 5 μM to 1.8 mM was performed using epithelial Madin-Darby canine kidney (MDCK) cells cultured alone or cocultured with rat bone marrow-derived MSC or preexposed to MSC-conditioned medium. TJ assembly was measured by assessing ZO-1 relocation to cell-cell contacts. Experiments were conducted using MDCK stably expressing short-hairpin-RNA (shRNA) against LKB1 or luciferase (LUC, as controls). Compound STO-609 (50 μM) was used as CaMKK inhibitor.Results. Following Ca2+switch, ZO-1 relocation and phosphorylation/activation of AMPK were significantly higher in MDCK/MSC compared to MDCK. No difference in AMPK phosphorylation was observed between LKB1-shRNA and Luc-shRNA MDCK following Ca2+switch. Conversely, incubation with STO-609 prior to Ca2+switch prevented AMPK phosphorylation and ZO-1 relocation. MSC-conditioned medium slightly but significantly increased AMPK activation and accelerated TJ-associated distribution of ZO-1 post Ca2+switch in comparison to regular medium.Conclusions. MSC modulate the assembly of epithelial TJ, via the CaMKK/AMPK pathway independently of LKB1.
- Subjects :
- 0301 basic medicine
lcsh:Internal medicine
Article Subject
biology
Tight junction
Chemistry
Kinase
[SDV]Life Sciences [q-bio]
Mesenchymal stem cell
AMPK
Cell Biology
030204 cardiovascular system & hematology
Cell biology
Small hairpin RNA
[SDV] Life Sciences [q-bio]
03 medical and health sciences
030104 developmental biology
0302 clinical medicine
AMP-activated protein kinase
biology.protein
Phosphorylation
lcsh:RC31-1245
Protein kinase A
Molecular Biology
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 1687966X and 16879678
- Database :
- OpenAIRE
- Journal :
- Stem Cells International, Stem Cells International, 2017, Stem Cells International, Hindawi Publishing Corporation, 2017, Stem Cells International, Vol 2017 (2017)
- Accession number :
- edsair.doi.dedup.....2a0f3aa449f57d4fc6c2d01a9ed4a472