Department of Orthopedic Surgery, Chongming Branched Hospital of Xinhua Hospital Affiliated to Medical School of Shanghai Jiaotong Universtiy, Shanghai 202150, China.
Department of Orthopedic Surgery, Chongming Branched Hospital of Xinhua Hospital Affiliated to Medical School of Shanghai Jiaotong Universtiy, Shanghai 202150, China.
Biomed Pharmacother. 2018 Mar;99:431-437. doi: 10.1016/j.biopha.2018.01.053.
Nucleus pulposus (NP) cell senescence correlates with disc degeneration. Previous studies imply that inflammation induces NP cell senescence. Osteogenic protein-1 (OP-1) is helpful for regenerating degenerative disc.
To investigate whether OP-1 treatment can protect against the inflammatory cytokine TNF-α-induced NP cell senescence, and its potential mechanism.
Rat NP cells were cultured in either basic culture medium or basic culture medium with the inflammatory cytokine TNF-α for 3 days. OP-1 was added into the culture medium of TNF-α-treated NP cells to investigate the protective effects of OP-1 against cellular senescence. NP cell senescence was evaluated by some direct and indirect parameters, such as senescence associated β-galactosidase (SA-β-Gal) activity, cell cycle, telomerase activity, expression of senescence markers (p16 and p53), and the matrix homeostatic phenotype. Additionally, intracellular reactive oxygen species (ROS) and the activity of the NF-κB pathway were also analyzed.
Compared with the control group, the inflammatory cytokine TNF-α significantly promoted NP cell senescence, reflected by the increase in SA-β-Gal activity, G0/G1 phase fraction and senescence markers (p16 and p53) expression, and the decrease in telomerase activity and matrix macromolecules (aggrecan and collagen II) expression. However, OP-1 suppressed the effects of TNF-α on NP cell senescence. Further analysis showed that OP-1 reduced ROS generation in TNF-α-treated NP cells, as well as the resulting activity of the NF-κB pathway.
OP-1 can attenuate inflammatory cytokine TNF-α-induced NP cell senescence, and the ROS/ NF-κB pathway may participate in this regulatory process. This study provides that OP-1 may be efficacy in retarding inflammation-exacerbated disc degeneration.
髓核（NP）细胞衰老与椎间盘退变相关。既往研究提示炎症可诱导 NP 细胞衰老。骨形成蛋白-1（OP-1）有助于退变椎间盘的再生。
探讨 OP-1 治疗是否可以预防炎症细胞因子 TNF-α诱导的 NP 细胞衰老及其潜在机制。
将大鼠 NP 细胞分别在基础培养基或含炎症细胞因子 TNF-α的基础培养基中培养 3 天。向 TNF-α处理的 NP 细胞培养基中加入 OP-1，以研究 OP-1 对细胞衰老的保护作用。通过衰老相关β-半乳糖苷酶（SA-β-Gal）活性、细胞周期、端粒酶活性、衰老标志物（p16 和 p53）表达及基质稳态表型等直接和间接参数评估 NP 细胞衰老。此外，还分析了细胞内活性氧（ROS）和 NF-κB 通路的活性。
与对照组相比，炎症细胞因子 TNF-α可显著促进 NP 细胞衰老，表现为 SA-β-Gal 活性增加、G0/G1 期细胞比例增加、衰老标志物（p16 和 p53）表达增加、端粒酶活性降低以及基质大分子（聚集蛋白聚糖和 II 型胶原）表达减少。然而，OP-1 抑制了 TNF-α对 NP 细胞衰老的作用。进一步分析表明，OP-1 减少了 TNF-α处理的 NP 细胞中的 ROS 生成，以及由此产生的 NF-κB 通路活性。
OP-1 可减轻炎症细胞因子 TNF-α诱导的 NP 细胞衰老，ROS/NF-κB 通路可能参与这一调节过程。本研究提示 OP-1 可能具有减缓炎症加重的椎间盘退变的作用。
