NF-κB信号通路参与高糖在椎间盘退行性变中影响及其作用的研究

目的:近来研究发现,椎间盘退变与代谢性疾病,尤其是与糖尿病具有明显的相关性,但具体机制尚未有深入研究。本实验拟探究高糖微环境诱导椎间盘退行性变及其对NF-κB信号通路的影响,为进一步揭示高糖诱导椎间盘髓核细胞退变的机制提供研究基础,为延缓、阻止糖尿病椎间盘退变和治疗糖尿病相关腰痛疾病带来新的策略和方法。方法:1、高糖微环境与IVDD的关系:使用5.5 mmol/L、15 mmol/L、30 mmol/L、100 mmol/L不同浓度葡萄糖培养基培养髓核细胞,RT-PCR检测髓核细胞MMP-3、MMP-13、Aggrecan、CollagenII的表达;2、NF-κB信号通路参与高糖微环境调控IVDD进展:Bay11-7082抑制NF-κB信号通路激活,再使用RT-PCR、Western Blot检测髓核细胞MMP-3、MMP-13、Aggrecan、CollagenII和NF-κB的表达。结果:RT-PCR检测显示,在不同葡萄糖浓度下,Aggrecan、CollagenII随浓度升高表达减少,MMP-3、MMP-13随浓度升高表达增加。RT-PCR、Western Blot检测显示,使用Bay11-7082可使高糖组中Aggrecan、CollagenII表达增加,MMP-3、MMP-13表达减少。结论:高糖微环境诱导椎间盘退行性变发病,且NF-κB信号通路参与高糖微环境诱导椎间盘退行性变发病。

The Study of High Glucose Promoting Intervertebral Disc Degeneration through NF-kappa B Signaling Pathway

ABSTRACT Objective: Recently, some researches show that intervertebral disc degeneration is significantly related to metabolic diseases, especially diabetes, but the specific mechanism has not been further studied. This study aims to the role of high glucose microenvironment and NF-kappa B signaling pathway in IVDD and to explore the mechanism of high glucose-induced degeneration of nucleus pulposus cells through NF-kappa B signaling pathway. It may provide a new strategy and method for preventing and treating diabetic-related low back pain. Methods: Nucleus pulposus cells was cultured in 5.5 mmol/L, 15 mmol/L, 30 mmol/L and 100 mmol/L glucose medium and the expression of MMP-3, MMP-13, Aggrecan and CollagenII in nucleus pulposus cells was detected by RT-PCR. Bay11-7082 was used to inhibit the activation of NF-kappa B signaling pathway and the expression of MMP-3, MMP-13, Aggrecan, CollagenII and NF-kappa B in nucleus pulposus cells was detected by RT-PCR and Western Blot. Results: RT-PCR showed that at different concentration, the expression of Aggrecan and CollagenII decreased with the increasing of concentration, while the expression of MMP-3 and MMP-13 increased with the increasing of concentration. RT-PCR and Western Blot showed that after using Bay11-7082, the expression of Aggrecan and CollagenII increased and the expression of MMP-3 and MMP-13 decreased in the high glucose group. Conclusion: High glucose microenvironment is correlated with intervertebral disc degeneration. High glucose microenvironment induces intervertebral disc degeneration by mediating NF-kappa B signaling pathway.