基于PTGFR-IP3/DAG-Ca2+通路研究莪术油倍半萜治疗痛经的物质基础及作用机制

痛经是当今世界尚未攻克的医学难题，严重危害广大女性健康。莪术为代表性活血化瘀药，治疗痛经疗效显著，以莪术油效果更佳，但其相关物质基础及机制研究匮乏。申请人前期发现莪术油倍半萜具有明显治疗痛经的作用，通过机制探索明确PTGFR为其重要靶点，作用通路与外钙内流和内钙释放有关。因此，提出科学假说：“倍半萜是莪术油治疗痛经的重要药效物质，其作用机制与PTGFR-IP3/DAG-Ca2+信号通路有关”。拟以“靶向分离-体外评价-体内验证-机制研究”模式开展研究：应用HPLC-HRMS-SPE-NMR等技术快速定向分离倍半萜成分；以离体子宫收缩模型和大鼠原发性痛经模型系统评价单体活性；利用激光共聚焦、膜片钳、Western blot、Real-time PCR等技术研究成分对PTGFR-IP3/DAG-Ca2+通路的影响，以期全面揭示莪术治疗痛经的物质基础及作用机制，为现代临床治疗妇科疾病提供新方式。

Dysmenorrhea is the unsolved medical problem in the world, which seriously endangers the health of women. Curcuma phaeocaulis, as a representative Chinese medicine for blood-activating and stasis-resolving, has a significant effect on treating dysmenorrhea. Especially, the essential oil of C. phaeocaulis shows more prominent therapeutic effect on dysmenorrhea. However, there are few studies on the material basis and the mechanism of it. The applicant previously discovered that sesquiterpenoids from the essential oil of C. phaeocaulis have obvious effects on the treatment of dysmenorrhea. Furthermore, through mechanism exploration, it was clear that PTGFR is the target of action, and the pathway of action is related to external calcium influx and internal calcium release. Therefore, the project puts forward an academic hypothesis that sesquiterpenoids are important pharmacodynamic substances of the essential oil of C. phaeocaulis for the treatment of dysmenorrhea, and the mechanism is related to the PTGFR-IP3/DAG-Ca2+ pathway. It is planned to conduct an in-depth research based on the model of "targeted separation-in vitro evaluation-in vivo verification-mechanism research". Firstly, the sesquiterpenoids are rapidly and directionally separated by using HPLC-HRMS-SPE-NMR and other techniques. Then, the isolated uterine contraction model and the rat primary dysmenorrhea model are applied to evaluate the effectiveness of the compounds. Eventually, the effects of the active constituents on the PTGFR-IP3/DAG-Ca2+ signaling pathway are explored by laser confocal, patch clamp, western blot, real-time PCR and other techniques. This project is hopeful to fully reveal the material basis and the molecular mechanism of C. phaeocaulis in treatment of dysmenorrhea, and provide a new way for modern clinical treatment of gynecological diseases.