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Journal of Korean Neurosurgical Society 2010;47(1): 30-35.
doi: https://doi.org/10.3340/jkns.2010.47.1.30
The Effects of TWEAK, Fn14, and TGF-beta1 on Degeneration of Human Intervertebral Disc.
Hoon Huh, Yong Jik Lee, Jung Hee Kim, Min Ho Kong, Kwan Young Song, Gun Choi
1Department of Neurosurgery, Medical Institute, Seoul Medical Center, Seoul, Korea.
2Division of Clinical Research, Medical Institute, Seoul Medical Center, Seoul, Korea. lyj2333@hanmail.net
3Clinical Division of Spine, Wooridul Spine Hospital, Seoul, Korea.
ABSTRACT
OBJECTIVE
The purpose of this study is to explain the effect and reciprocal action among tumor necrosis factor (TNF) like weak inducer of apoptosis (TWEAK), fibroblast growth factor-inducible 14 (Fn14), and transforming growth factor-beta1 (TGF-beta1) on degeneration of human intervertebral disc (IVD).
METHODS
Human intervertebral disc tissues and cells were cultured with Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM/F-12) media in 37degrees C, 5% CO2 incubator. When IVD tissues were cultured with TWEAK, Fn14 that is an antagonistic receptor for TWEAK and TGF-beta1, the level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay and sex determining region Y (SRY)-box 9 (Sox9) and versican messenger ribonucleic acid (mRNA) levels were estimated by reverse transcriptase polymerase chain reaction (RT-PCR).
RESULTS
When human IVD tissue was cultured for nine days, the sGAG content was elevated in proportion to culture duration. The sGAG was decreased significantly by TWEAK 100 ng/mL, however, Fn14 500 ng/mL did not change the sGAG production of IVD tissue. The Fn14 increased versican and Sox9 mRNA levels decreased with TWEAK in IVD tissue TGF-beta1 20 ng/mL elevated the sGAG concentration 40% more than control. The sGAG amount decreased with TWEAK was increased with Fn14 or TGF-beta1 but the result was insignificant statistically. TGF-beta1 increased the Sox9 mRNA expression to 180% compared to control group in IVD tissue. Sox9 and versican mRNA levels decreased by TWEAK were increased with TGF-beta1 in primary cultured IVD cells, however, Fn14 did not show increasing effect on Sox9 and versican.
CONCLUSION
This study suggests that TWEAK would act a role in intervertebral disc degeneration through decreasing sGAG and the mRNA level of versican and Sox9.
Key Words: Intervertebral disc; TWEAK; Fn14; TGF-beta1; Sox9; Versican
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