Role of nuclear factor of activated T-cells in regulating ADAMTS-4 promoter activity in nucleus pulposus cells
10.3724/SP.J.1008.2014.00495
- Author:
Peng CAO
1
Author Information
1. Department of Orthopedic Surgery, Changzheng Hospital, Second Military Medical University
- Publication Type:Journal Article
- Keywords:
ADAMTS-4;
Intervertebral disk;
Nuclear factor of activated T-cells;
Nucleus pulposus cells
- From:
Academic Journal of Second Military Medical University
2014;35(5):495-499
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To observe the regulatory effect of nuclear factor of activated T-cells (NFAT)-1, 4, 5 on ADAMTS-4. promoter activity in nucleus pulposus cells, so as to discuss the underlying molecular mechanism of intervertebral disc degeneration. Methods: The ADAMTS-4-pβ-gal-Basic was inserted into PGL3-Basic vector after double digestion by the restriction enzyme Xho I and Hind III. The purified PGL3-ADAMTS-4 plasmid was obtained through screening. Rat nucleus pulposus cells were cultured in vitro and, via Lipofectamine2000 transfection reagent, transfected with NFAT-1, NFAT-4, NFAT-5 or DN-TonEBP expressing plasmids with or without appropriate backbone vector and 175 ng ADAMTS-4 promoter. The normal cells were taken as controls and transfected cells were taken as treatment group. Dual-Luciferase™ reporter assay system was used to detect the effect of NFAT-1, 4, 5(TonEBP) on ADAMTS-4 promoter activity. Results: We successfully constructed PGL3-ADAMTS-4 plasmid and identified two NFAT-Bind elements in the promoter. Compared with the control group, ADAMTS-4 promoter activity was significantly inhibited in NFAT-1 group (P < 0. 05). While the activities of ADAMTS-4 promoter in NFAT-4, NAFT-5 and DN-TonEBP groups showed no significant changes(P>0. 05). Conclusion: NFAT-1 can regulate ADAMTS-4 expression, which may play a role in modulating aggrecan content in the intervertebral disc physiology and/or intervertebral disc degeneration, providing a new strategy for biological treatment of intervertebral disc degeneration.
