17β-雌二醇负调控NF-κB-MMP-3/13信号通路抑制椎间盘退变的作用机制研究

Clinically, spinal degenerative diseases are commonly seen and intervertebral disc degeneration (IVDD) is known as the main pathological basis of those diseases. Our previous studies have revealed that 17beta-estradiol (E2) is able to protect intervertebral disc cells from apoptosis as well as inhibit IVDD by downregulating the expression of MMP-3/13; all the effects of E2 can be abolished by the use of ICI182780, an estrogen receptor (ER) antagonist. Therefore, a hypothesis develops to be that E2 may inhibit IVDD by negatively regulating NF-kappa B-MMP-3/13 signaling pathway. The objective of this study is to elucidate whether E2 inhibits IVDD by negatively regulating NF-kappa B-MMP-3/13 signaling pathway through a variety of experimental methods including IHC, Western blot, qPCR, and CHIP; both in-vivo and in-vitro analyses will be performed and some of them are based on rat OVX models and rat IVDD models. Furthermore, inhibitors of the signaling pathway as well as RNAi will be used to reversely clarify the mechanism above. Taken together, our work is expected to provide theoretical basis for future use of E2 to prevent or treat IVDD-related diseases in a clinical situation.

脊柱退行性疾病是临床常见病，椎间盘退变是其主要病理基础。本课题组前期研究表明，17β-雌二醇（E2）可有效抑制椎间盘细胞的凋亡，通过下调基质金属蛋白酶MMP-3/13的表达发挥椎间盘保护作用；而应用雌激素受体（ER）抑制剂ICI182780后，E2对MMP-3/13的负调控作用消失且不能有效抑制椎间盘细胞的异常凋亡并丧失对椎间盘的保护作用。因此，本课题组提出了E2负调控NF-κB-MMP-3/13信号通路抑制椎间盘退变的假说。本项目应用大鼠OVX模型和尾椎间盘退变模型，采用IHC、Western blot、qPCR、CHIP等多种实验技术，从在体和离体两个水平，分析NF-κB-MMP-3/13信号通路是否在E2负调控MMP-3/13抑制椎间盘退变中发挥关键作用，并应用信号通路抑制剂和RNAi从反面进一步验证。本研究为以后临床转化应用E2防治椎间盘退变性疾病提供理论依据。

脊柱退行性疾病是临床常见病，椎间盘退变是其主要病理基础。国内外研究和我们的前期研究均表明，17β-雌二醇（E2）可有效抑制椎间盘细胞的凋亡，通过下调基质金属蛋白酶MMP-3/13的表达发挥椎间盘保护作用。因此，本研究提出了E2负调控NF-κB-MMP-3/13信号通路抑制椎间盘退变的假说。本项目通过针刺联合卵巢切除构建了大鼠尾椎间盘退变模型，并利用这一模型验证17β-雌二醇在体水平对椎间盘退变的保护作用。在离体水平和在体水平，本项目验证了E2-NF-κB-MMP-3/13信号通路在抑制椎间盘退变过程中的作用，并分析了该信号通路与椎间盘退变的临床相关性。本研究的结果显示，椎间盘退变过程中NF-κB信号通路激活，促进了细胞外基质降解；而雌激素通过与其受体Erβ结合抑制NF-κB信号通路的激活，下调了MMP-3/13的表达，抑制了细胞外基质降解，发挥了对椎间盘的保护作用。本项目相关研究成果已发表4篇SCI论文，包括2篇论著和2篇综述，其中1篇综述被收录为高被引论文。本课题研究结果旨在为将来应用雌激素延缓椎间盘退变进程提供一定的理论依据，从而更好的造福于广大椎间盘疾病患者。