基于Fas/FasL信号通路回医烙灸对腰椎间盘突出后重吸收作用机制的研究

The Resorption of Herniated Lumbar Discs（RHLD）bring the new enlightenment for conservative treatment to cure lumbar disc herniation.How to promote RHLD is one of the key problems that need to solve conservative treatment.In Hui medicine, cautery moxibustion is one of the most effective treatments of lumbar disc herniation.The cautery moxibustion reduce the recurrence rate of lumbar disc herniation,but the mechanism of cautery moxibustion to treat RHLD can be expound. Apoptosis of nucleus pulposus cells is the major cause of RHLD.In our previous studies on rats, we have found that cautery moxibustion therapy can promote the apoptosis of nucleus pulposus cells,Caspase-3 and the IL-1 in the herniated lumbar discs.While IL-1 and Caspase-3 are key factors of the Fas/FasL signal pathways,and activating this pathway can increase the apoptosis of cartilage cells. In the present study，through analyzing the regulation of Fas/FasL signal pathway to the apoptosis of nucleus pulposus cells and the extracellular matrix of nucleus pulposus cells in herniated lumbar discs. Therefore the hypothesis that the cautery moxibustion possibly treats RHLD through mediating the Fas/FasL signaling pathway is proposed.In this study the model of cautery moxibustion intervention of rats’RHLD is adopted. Research contents:1. The mechanism of the cautery moxibustion influence multiplication and apoptosis of nucleus pulposus cells through mediating the Fas/FasL signaling pathway;2. The mechanism of the cautery moxibustion regulate the extracellular matrix of nucleus pulposus cells through mediating the Fas/FasL signaling pathway.The purpose of the study is to testify the hypothesis that the cautery moxibustion may treat RHLD through mediating Fas/FasL signaling pathway.The study lay the foundation for promotion of the cautery moxibustion.

腰椎间盘突出后重吸收(RHLD)理论为保守疗法防治腰椎间盘突出症带来新的启示。特色回医烙灸治疗腰椎间盘突出症临床疗效确切，并能促进部分RHLD，降低腰椎间盘突出症的复发率，但作用机制不明。突出的髓核细胞凋亡在RHLD发挥重要作用。我们前期研究发现：烙灸可增加髓核细胞凋亡及髓核组织中IL-1和Caspase-3的表达。IL-1和Caspase-3是Fas/FasL信号通路上下游关键作用因子，激活该信号通路可促使髓核细胞凋亡，提出烙灸可能是介导Fas/FasL信号通路促进腰椎间盘突出后髓核重吸收的假说。本课题采用烙灸干预大鼠RHLD模型，主要研究：1.烙灸通过Fas/FasL通路对髓核细胞凋亡的调节机制；2.烙灸通过Fas/FasL通路对髓核细胞外基质的调控机制。验证烙灸介导Fas/FasL信号通路促进腰椎间盘突出后髓核重吸收的假说，阐明其作用机制，为烙灸防治腰椎间盘突出症提供理论和实验根据。

1.项目背景. 前期实验发现：回医烙灸疗法不仅可以促进大鼠RHLD模型椎间盘的重吸收，促使突出髓核细胞的凋亡，还可以导致髓核组织中IL-1和Caspase-3的高表达。IL-1和Caspase-3是Fas/FasL信号通路上下游的关键作用因子，同时激活Fas/FasL信号通路可增加髓核细胞的凋亡。由此我们提出回医烙灸疗法可能是通过介导Fas/FasL信号通路促进腰椎间盘突出后髓核重吸收的假说。在回医烙灸疗法促进突出间盘重吸收和减少腰椎间盘突出症复发率的基础上，进一步探讨其对突出椎间盘重吸收的作用机制具有重要意义。.2.主要研究内容. ① 分析回医烙灸疗法干预后实验大鼠行为学、影像学和髓核形态学评分的变化与Fas和FasL蛋白表达的相关性，为验证烙灸疗法通过介导Fas/FasL信号通路提供依据；. ② 探讨回医烙灸疗法通过Fas/FasL信号通路对髓核细胞凋亡的调节机制； ③ 研究回医烙灸疗法介导Fas/FasL信号通路对髓核细胞外基质环境的调控机制； ④ 采用阻滞剂抗Fas抗体抑制Fas/FasL信号通路，研究回医烙灸疗法对髓核细胞凋亡和细胞基质合成代谢平衡的影响。通过上述实验观察和检测，探讨回医烙灸疗法促进腰椎间盘突出后髓核重吸收作用机制。.3.重要结果.①回医烙灸疗法促进破裂腰椎间盘突出的重吸收。 ②回医烙灸疗法可以明显改善大鼠髓核重吸收模型的行为学。③回医烙灸疗法可以促进大鼠椎间盘髓核细胞的凋亡，而调节Fas/FasL信号通路也可促使髓核细胞的凋亡。④回医烙灸疗法可以增加髓核组织中IL-1和Caspase-3的表达。 .4.关键数据. ①发生髓核重吸收患者的腰椎间盘突出症复发次数低于没有发生髓核重吸收的患者（P<0.05）。②干预后回医烙灸组的运动功能评分明显低于其干预前的评分（P<0.01），干预后回医烙灸组的械痛阈触觉阈值改变百分率明显高于其干预前的评分（P<0.01）。③回医烙灸组和模型对照组的髓核细胞凋亡指数明显高于正常对照组（P<0.05），而回医烙灸组的髓核细胞凋亡指数高于模型对照组（P<0.05）。④回医烙灸组IL-1和Caspase-3的阳性细胞率高于模型对照组和空白对照组（P<0.05、P<0.01）。