Upregulated functional expression of Toll like receptor 4 in mesenchymal stem cells induced by lipopolysaccharide.
BACKGROUND
The coordinated change of haematopoietic supporting microenvironment in bone marrow (BM) is crucial for innate immunity and inflammation. As the precursors of marrow stroma, BM derived mesenchymal stem cells (MSCs) promote haematopoietic function, but their roles in innate immunity or inflammation have not been investigated. Here we investigated the expression of Toll like receptor 4 (TLR-4) and the effect of lipopolysaccharide (LPS) on its expression in BM MSCs in vitro.


METHODS
MSCs were harvested from adult rat's BM cells by density gradient centrifugation and adhesive culture. The purity of MSCs were identified with the cell morphological feature and osteogenic capacity, the phenotypes were tested by flow cytometry. Cultured MSCs were treated by LPS (1 microg/ml, 10 microg/ml or 100 microg/ml) for 24 hours. The relative expression levels of TLR-4 mRNA were detected by semiquantitative reverse transcription polymerase chain reaction and costimulatory molecules (CD80, CD86 and MHC-II) expressed on MSCs were analyzed by flow cytometry. The levels of tumor necrosis factor-alpha (TNF-alpha) in supernatants were determined by enzyme linked immunosorbent assay.


RESULTS
After incubation with LPS, MSCs expressed the higher levels of TLR-4 mRNA, costimulatory molecules and TNF-alpha than the untreated group: LPS 10 microg/ml was the most effective (P < 0.01); the levels of TLR-4 mRNA, costimulatory molecules and TNF-alpha decreased when MSCs were exposed to 100 microg/ml LPS. Except for MHC-II and TNF-alpha (P > 0.05), the levels of CD80, CD86 and TLR-4 mRNA were significantly lower than that in the treated group of 10 microg/ml (P < 0.01).


CONCLUSIONS
MSCs expressed TLR-4 mRNA. LPS activated the functional expression levels of TLR-4 in MSCs although the activity may depend on the concentration of LPS.
Shi
Liang
L
Department of Hematology, Affiliated Hospital of Guiyang Medical College, Guiyang 550004, China.
Wang
Ji-shi
JS
Liu
Xing-mei
XM
Hu
Xiao-yan
XY
Fang
Qin
Q
D000818:Animals; D051940:B7-2 Antigen / Q000032:analysis; D001854:Bone Marrow Cells / Q000276:immunology; D002454:Cell Differentiation; D002478:Cells, Cultured; D016130:Immunophenotyping; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000276:immunology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D051197:Toll-Like Receptor 4 / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D015854:Up-Regulation
D051940:B7-2 Antigen; D008070:Lipopolysaccharides; C495347:Tlr4 protein, rat; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha