Regulation and Signaling of TGF-β1 on the plasminogen activation system of human dental pulp cells
Date Issued
2015
Date
2015
Author(s)
Huang, Yu-An
Abstract
Aim: Transforming growth factor-β1 (TGF-β1) plays a role in the repair and dentinogenesis of human dental pulp, including the deposition and remodeling of extracellular matrix (ECM). Plasminogen activation system regulates ECM degradation. However the relationship between TGF-β1 and plasminogen activation system is not fully clear. The purpose of this study was to investigate the effect of TGF-β1 on plasminogen activation system of human dental pulp cells and its related signaling pathways. Materials and methods: Primary human dental pulp cells were treated with 0.5, 1, 5, 10, 25 ng/mL of TGF-β1. MTT assay, Reverse Transcription Polymerase Chain Reaction) (RT-PCR), Western Blot and Immunofluorescence were used to detect the effect of TGF-β1 on cell viability and expression of urokinase-type plasminogen activator (uPA), urokinase-type plasminogen activator receptor (uPAR), plasminogen activator inhibitor-1 (PAI-1) and collagen I at 24 hrs and 5 days. The activation of Smad2, TAK1, and ERK was also addressed. Besides, cells were pretreated with SB431542 (an ALK5/Smad2/3 inhibitor), 5z-7-oxozeaenol (a TAK1 inhibitor), U0126 (a MEK/ERK inhibitor) for examining the related signaling pathways. Results: TGF-β1 slightly inhibited cell proliferation in a dose-dependent manner, which could be reversed by SB431542 but not be reversed by U0126, and the cell number decreased more when adding 5z-7-oxozeaenol. TGF-β1 up-regulated PAI-1 and uPAR expression, whereas uPA was down-regulated and the changes of collagen I was not obvious. SB431542, 5z-7-oxozeaenol and U0126 generally could reverse the expression of TGF-β1 on PAI-1/uPAR; besides, SB431542 and 5z-7-oxozeaenol might have influence on uPA expression. Furthermore, TGF-β1 induced the activation of p-Smad2, p-Smad3, p-TAK1, and p-ERK signaling. Conclusion: The regulation of signaling pathway on TGF-β1-treated dental pulp cells is complicated. TGF-β1 can affect PAI-1/uPA/uPAR expression through Smad/ALK5 and Non-Smad (TAK1 and MEK/ERK) pathways and therefore affect the deposition and remodeling of ECM. These results give an aspect to understand the mechanism of pulpal repair, and can be helpful for further investigation of pulpo-dentin regeneration.
Subjects
human dental pulp cell
TGF-β1
PAI-1
uPA
uPAR
Type
thesis
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