MAPK/CREB信号转导通路与子宫内膜异位症疼痛敏化调控机制研究

Pain is the key symptom in endometriosis, but the efficacy of the current treatments including medical and surgical therapy is still not satisfactory. Recent studies have found that endometriosis patients experience neuropathic pain, through peripheral sensitization in the primary nociceptive neurons, and, central sensitization in the dorsal horn of the spinal cord. The precise mechanisms of sustained, chronic pain in endometriosis are unclear though MAPK/CREB signaling pathway is believed to play an important role in mechanisms of pain regulation. ..In this project, we will use the SD rat model of endometriosis to measure changes associated with pain in vivo, together with cell culture of dorsal root ganglion cells in vitro. .(1).to create and evaluate gross behavioural changes associated with painful stimuli,.(2).to study immunohistochemical changes in dorsal root ganglia and endometriosis lesions using a panel of antibodies (anti-MAPK, anti-CREB, anti-NGF, etc),.(3).to use Western blotting to identify changes in protein levels in dorsal root ganglia and endometriosis lesions associated with chronic pain...At the same time, a culture of dorsal root ganglion cells will be performed to measure the effects of different interventions to identify key molecular proteins in neural signal transduction of endometriosis-associated pain...These studies will provide insights into the mechanisms of pain in this animal model of endometriosis.

子宫内膜异位症（内异症）是一种严重影响妇女健康的常见病，发病率逐年上升。疼痛是内异症治疗的关键及难题，目前治疗内异症疼痛的药物或手术方法均不令人满意，因此探索更优的内异症疼痛治疗方法迫在眉睫。最新研究发现，神经病理性疼痛，包括外周敏化和中枢敏化，在内异症患者长期持续性的慢性疼痛发生中发挥重要作用，但机制不清。MAPK/CREB信号转导通路可能是内异症疼痛敏化调控的重要机制。本项目拟应用内异症SD大鼠模型，采用行为学、免疫组织化学和免疫印迹方法测定内异症不同干预前后DRG神经元和脊髓背角神经元MAPK/CREB信号通路变化，及其与异位病灶神经分布、模型疼痛行为改变的关系；应用离体DRG神经元细胞培养模型，研究雌激素及内异症相关活性因子干预前后MAPK/CREB信号通路变化及调控机制，以筛检MAPK/CREB信号转导通路介导内异症疼痛的关键分子，为内异症疼痛治疗提供更有效的治疗新靶点。

子宫内膜异位症（内异症）疼痛的发生机制目前尚不完全清楚，近年研究发现，内异症疼痛既是一种炎症性疼痛又是一种伤害性与神经源性疼痛，MAPK是内异症炎症介质的主要调控者。因而本项目设计研究MAPK/CREB信号通路在内异症外周与中枢疼痛敏化中的作用。研究发现，内异症病灶以及脊髓背角和DRG神经元细胞均存在MAPK信号通路分子MAPK、CREB、BDNF及NGF水平的高表达，其表达水平与内异症模型SD大鼠热痛觉、神经纤维密度以及患者疼痛相关。雌激素在体干预模型SD大鼠可使模型病灶与DRG 组织中的MAPK信号通路分子MAPK、CREB、BDNF及NGF的表达水平显著增加，而给予不同抑制剂在体干预模型SD大鼠可使模型病灶与DRG 组织中的MAPK信号通路分子MAPK、CREB、BDNF及NGF的表达水平显著下降。模型离体DRG细胞与内异症病灶异位内膜间质细胞给予雌激素干预可使其MAPK信号通路分子MAPK、CREB、BDNF及NGF的表达水平迅速增加，而给予不同抑制剂干预可使其MAPK信号通路分子MAPK、CREB、BDNF及NGF的表达水平迅速下降。离体干预实验与在体干预实验得出几乎相同的结论。由此可见，雌激素是内异症发病及疼痛触发的关键始动促发因素，MAPK信号通路介导雌激素在内异症疼痛敏化中发挥关键作用，选择合适关键分子尤其是MAPK抑制剂给予MAPK信号通路干预可为内异症疼痛提供的治疗新靶点。