PMID- 29483719
OWN - NLM
STAT- MEDLINE
DCOM- 20190304
LR  - 20190304
IS  - 1671-167X (Print)
IS  - 1671-167X (Linking)
VI  - 50
IP  - 1
DP  - 2018 Feb 18
TI  - [Decreased phosphorylation of mitogen activated protein kinase and protein kinase 
      B contribute to the inhibition of osteogenic differentiation mediated by 
      activation of Toll like receptor in human periodontal ligament stem cells].
PG  - 33-41
AB  - OBJECTIVE: To investigate the effects of Toll like receptors on the osteogenesis 
      of human pe-riodontal ligament stem cells (hPDLSCs) and probable molecular 
      mechanism. METHODS: Real-time PCR and flow cytometry were applied to test the 
      expression of TLRs in hPDLSCs and the positive cell percentage of TLR. hPDLSCs 
      were cultured in osteogenic medium for 7 to 14 days with different TLR agonists 
      at various concentrations . The effect of different TLR on osteogenic 
      differentiation of hPDLSCs was evaluated by alizarin red S staining, alkaline 
      phosphatase (ALP) staining and ALP activity assay. Western blotting was used to 
      analyze the phosphorylation levels of extracellular regulated protein kinases 
      (ERK), c-Jun N-terminal protein kinase (JNK), P38, AKT and expression of Runx2 an 
      osteogenic related gene after treatment with TLR agonists, compared with the 
      effect of inhibitors of mitogen activated protein kinase (MAPK) or protein kinase 
      B (PKB or AKT) on Runx2 expression of hPDLSCs cultured in osteogenic medium. 
      RESULTS: Higher expressions of TLR1,3,4,6 were found in hPDLSCs through real-time 
      PCR. Positive cell percentage of TLR was determined by flow cytometry and 
      described as TLR1: 2.82%+/-0.68%; TLR2: 1.26%+/-0.09%; TLR3: 13.23%+/-2.05%; TLR4: 
      3.64%+/-0.79%; TLR6: 3.21%+/-1.64%, whose tendency was comparable to their mRNA 
      expression in hPDLSCs. Most TLR ligands had no effect on the ALP staining, 
      activity and mineralization of hPDLSCs at lower concentration except for 0.1 mg/L 
      PolyI:C could induce the osteogenic ability of hPDLSCs. On the contrary, Higher 
      concentration of TLR ligands (PolyI:C: 10 mg/L, LPS: 10 mg/L , Pam3CSK4: 1 mg/L, 
      FSL-1: 50 mug/L) had obviously inhibitory effect on osteogenic differentiation of 
      hPDLSCs. Activation of TLR using higher concentration of TLR ligands could 
      downregulate the phosphorylation levels of ERK, P38, JNK and AKT, and also 
      reduced the expression of Runx2, compared with the untreated control. The 
      inhibitors of MAPK (U0126, SP600125,SB203580) and inhibitor of AKT (perifosine) 
      could also inhibit Runx2 expression. CONCLUSION: Higher concentration of TLR 
      ligands could inhibit osteogenic differentiation of hPDLSCs. This inhibitory 
      effect seemed to be related to decreased phosphorylation of MAPK and AKT.
FAU - Zhu, Y Y
AU  - Zhu YY
AD  - Department of Orthodontics, Peking University School and Hospital of Stomatology 
      & National Engineering Laboratory for Digital and Material Technology of 
      Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, 
      China.
FAU - Li, Q
AU  - Li Q
AD  - Department of Orthodontics, Peking University School and Hospital of Stomatology 
      & National Engineering Laboratory for Digital and Material Technology of 
      Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, 
      China.
FAU - Zhang, Y M
AU  - Zhang YM
AD  - Department of Orthodontics, Peking University School and Hospital of Stomatology 
      & National Engineering Laboratory for Digital and Material Technology of 
      Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, 
      China.
FAU - Zhou, Y H
AU  - Zhou YH
AD  - Department of Orthodontics, Peking University School and Hospital of Stomatology 
      & National Engineering Laboratory for Digital and Material Technology of 
      Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, 
      China.
LA  - chi
PT  - Journal Article
PL  - China
TA  - Beijing Da Xue Xue Bao Yi Xue Ban
JT  - Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health 
      sciences
JID - 101125284
RN  - 0 (Toll-Like Receptors)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
SB  - IM
MH  - *Cell Differentiation
MH  - Cells, Cultured
MH  - Humans
MH  - Ligaments
MH  - *Mitogen-Activated Protein Kinases/metabolism
MH  - *Osteogenesis
MH  - *Periodontal Ligament/cytology/metabolism
MH  - Phosphorylation
MH  - *Proto-Oncogene Proteins c-akt/metabolism
MH  - Stem Cells
MH  - *Toll-Like Receptors/metabolism
EDAT- 2018/02/28 06:00
MHDA- 2019/03/05 06:00
CRDT- 2018/02/28 06:00
PHST- 2018/02/28 06:00 [entrez]
PHST- 2018/02/28 06:00 [pubmed]
PHST- 2019/03/05 06:00 [medline]
PST - ppublish
SO  - Beijing Da Xue Xue Bao Yi Xue Ban. 2018 Feb 18;50(1):33-41.