MyD88-dependent Toll-like receptor 4 signal pathway in intervertebral disc degeneration

Qin,Chuqiang; Zhang,Bo; Zhang,Liang; Zhang,Zhi; Wang,Le; Tang,Long; Li,Shuangqing; Yang,Yixi; Yang,Fuguo; Zhang,Ping; Yang,Bo

doi:10.3892/etm.2016.3425

MyD88-dependent Toll-like receptor 4 signal pathway in intervertebral disc degeneration

Authors:
- Chuqiang Qin
- Bo Zhang
- Liang Zhang
- Zhi Zhang
- Le Wang
- Long Tang
- Shuangqing Li
- Yixi Yang
- Fuguo Yang
- Ping Zhang
- Bo Yang
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Affiliations: Department of Orthopedic Surgery, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, Guangdong 510150, P.R. China
Published online on: June 6, 2016 https://doi.org/10.3892/etm.2016.3425
Pages: 611-618
Copyright: © Qin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Lower back pain (LBP) is a common and remitting problem. One of the primary causes of LBP is thought to be degeneration of the intervertebral disc (IVD). The aim of the present study was to investigate the role of the myeloid differentiation primary-response protein 88 (MyD88)‑dependent Toll‑like receptor 4 (TLR4) signal pathway in the mechanism of IVD degeneration. IVD nucleus pulposus cells isolated and cultured from the lumbar vertebrae of Wistar rats were stimulated by various doses of lipopolysaccharide (LPS; 0.1, 1, 10 and 100 µg/ml) to simulate IVD degeneration. Cells were rinsed and cultured in serum-free Dulbecco's modified Eagle's medium/F12. Reverse transcription-quantitative polymerase chain reaction was used to determine the levels of TLR4, MyD88, tumor necrosis factor α (TNFα), and interleukin‑1β (IL‑1β) mRNA expression after 1, 3, 6, 9 and 12 h of incubation. Additionally, western blot and enzyme‑linked immunosorbent assay analyses were used to determine the levels of TLR4, MyD88, TNFα, and IL‑1β protein expression after 24, 48 and 72 h of incubation. The levels of TLR4, MyD88, TNFα and IL‑1β mRNA all increased in the cells stimulated by 10 µg/ml LPS at 3, 6 and 9 h (all P<0.001). Furthermore, the levels of TLR4, MyD88, TNFα and IL‑1β protein all increased at 24, 48 and 72 h (all P<0.001). Additionally, the mRNA and protein levels of TLR4, MyD88, TNFα and IL‑1β increased significantly in the cells stimulated by 1, 10 and 100 µg/ml LPS compared with the control group, and reached a peak in the 10 µg/ml LPS group (all P<0.001). These results suggest that the MyD88-dependent TLR4 signal pathway is a target pathway in IVD degeneration. This pathway is time phase‑ and dose-dependent, and when activated can lead to the release of inflammatory factors that participate in IVD degeneration.

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