基于NGF/MAPK通路研究桂枝茯苓丸抑制子宫内膜异位症疼痛的机制

Endometriosis pain is neuropathic pain associated with inflammation. Western medicine has no long-term effective treatment. Guizhi Fuling capsule is commonly used in clinical treatment of endometriosis pain for a long time. Our preliminary clinical research proved that Guizhi Fuling capsule can relieve primary dysmenorrhea; animal experiments also confirmed that Guizhi Fuling capsule can not only significantly increased the pain threshold of endometriosis in rats, but also promote apoptosis of ectopic endometrial cells. However, the mechanism of Guizhi Fuling capsule inhibiting endometriosis pain is unclear. Recent research suggests that NGF/ MAPK signaling pathway is one of the important targets involved with pain. This project intends to establish a rat model of endometriosis to evaluate the efficacy of Guizhi Fuling capsule suppressing pain by detecting the change of the behavior of pain, inflammatory factors, nerve factors and receptor expression in the animal level; separate primary ectopic endometrial cells, DRG and dorsal horn neurons, to detect the expression level of the key factors in NGF/MAPK signaling pathway; using Guizhi Fuling capsule containing serum to intervene the ectopic endometrial cells to detect the expression change of the key factors in this pathway, and then clarify the mechanism of Guizhi Fuling capsule inhibiting endometriosis pain. The successful implementation of this project will contribute to the understanding of treatment of endometriosis pain by Guizhi Fuling capsule, meanwhile provide new theories and methods for finding drug targets suppressing pain.

子宫内膜异位症疼痛是一种伴随炎性痛的神经病理性疼痛，西医尚无有效的治疗药物。桂枝茯苓丸是治疗子宫内膜异位症的经典方剂。本课题组在临床研究中证实，桂枝茯苓丸具有抑制原发性痛经的作用；动物实验表明，桂枝茯苓丸显著提高异位症大鼠的疼痛阈值，促进异位内膜细胞的凋亡，但其抑制异位症疼痛的机理尚不明确。近期研究证明，NGF/MAPK信号通路是控制疼痛的重要靶点。本项目拟建立子宫内膜异位症模型，通过疼痛行为学、炎性因子、神经因子及受体表达的改变，从动物水平评价桂枝茯苓丸抑制疼痛的疗效；分离原代异位内膜细胞，DRG及脊髓背角神经元，测定NGF/MAPK信号通路关键因子的表达水平；用桂枝茯苓丸含药血清对异位内膜细胞进行干预，从分子水平研究此通路中关键因子表达水平的变化，进而阐明桂枝茯苓丸镇痛作用机理。本项目的顺利实施将会促进对桂枝茯苓丸治疗异位症疼痛的现代认识，同时为寻找抑制疼痛药物靶点提供新的理论和方法。

子宫内膜异位症是一种困扰女性健康的常见妇科疾病，其主诉症状是疼痛，西医尚无有效的治疗药物。桂枝茯苓丸出自东汉末年著名医学家张仲景的《金匮要略》，至今使用已有1800多年的历史，具有活血化瘀消癥之功效，是治疗子宫内膜异位症的经典方剂。本项目主要通过构建子宫内膜异位症大鼠动物模型，评价和探讨了经典处方“桂枝茯苓丸”在子宫内膜异位症的作用效果和作用机制。本研究发现，桂枝茯苓丸能明显减小大鼠异位组织的体积，提高子宫内膜异位症大鼠的疼痛阈值，缓解疾病引起的疼痛。深入的机制研究发现，桂枝茯苓丸主要通过改善子宫内膜异位引起的神经损伤、抑制神经营养因子的表达来发挥作用。其次，桂枝茯苓丸能显著抑制IL6、IL-1β等炎性因子的水平，进而抑制IL6/JAK-STAT3、MAPK/ERK信号通路，从而发挥缓解炎性痛的作用。此外，初步的临床研究发现，桂枝茯苓丸能有效改善原发性痛经，疼痛强度显著改善，疼痛持续时间明显降低。综上，本项目阐释了桂枝茯苓丸在子宫内膜异位症治疗中的药效和机制，为桂枝茯苓丸的临床应用提供了理论基础和科学依据。