Objectives. Evodia rutaecarpa (ER) is a well-known herbal Chinese medicine traditionally used for analgesia in dysmenorrhea, headaches, abdominal pain, etc. Notably, the analgesic effect of wine-processed Evodia rutaecarpa (PER) was more potent than that of raw ER. This research aimed to investigate the mechanism and pharmacodynamic substance basis of raw ER and PER on smooth muscle cells of dysmenorrhea mice. Methods. Metabolomics methods based on UPLC-Q-TOF-MS were utilized to analyse the differential components of ER before and after wine processing. Afterwards, the uterine smooth muscle cells were isolated from the uterine tissue of dysmenorrhea and normal mice. The isolated dysmenorrhea uterine smooth muscle cells were randomly divided into four groups: model group, 7-hydroxycoumarin group (1 mmol/L), chlorogenic acid (1 mmol/L), and limonin (50 μmol/L). The normal group consisted of the isolated normal mouse uterine smooth muscle cells, which were repeated 3 times in each group. The cell contraction and the expression of P2X3 and Ca²⁺ in vitro were determined using immunofluorescence staining and laser confocal; ELISA was used for detection of PGE2, ET-1, and NO content after 7-hydroxycoumarin, chlorogenic acid, and limonin administered for 24 h. Results. The metabolomics results suggested that seven differential compounds were identified in the extracts of raw ER and PER, including chlorogenic acid, 7-hydroxycoumarin, hydroxy evodiamine, laudanosine, evollionines A, limonin, and 1-methyl-2-[(z)-4-nonenyl]-4 (1H)-quinolone. The in vitro results showed that 7-hydroxycoumarin, chlorogenic acid, and limonin were able to inhibit cell contraction and PGE2, ET-1, P2X3, and Ca²⁺ in dysmenorrhea mouse uterine smooth muscle cells and increase the content of NO. Conclusion. Our finding suggested that the compounds of the PER were different from those of the raw ER, and 7-hydroxycoumarin, chlorogenic acid, and limonin could improve dysmenorrhea in mice whose uterine smooth muscle cell contraction was closed with endocrine factors and P2X3-Ca²⁺ pathway.

中文翻译：

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吴茱萸生酒酒对痛经小鼠平滑肌细胞的作用机制及药效物质基础

目标。吴茱萸（ER）是一种著名的中药，传统上用于镇痛痛经、头痛、腹痛等。值得注意的是，酒制吴茱萸（PER）的镇痛作用比生吴茱萸的镇痛效果更有效。本研究旨在探讨生ER和PER对痛经小鼠平滑肌细胞的作用机制及药效物质基础。方法。采用基于UPLC-Q-TOF-MS的代谢组学方法分析葡萄酒加工前后ER的差异成分。随后从痛经小鼠和正常小鼠的子宫组织中分离出子宫平滑肌细胞。将分离的痛经子宫平滑肌细胞随机分为四组：模型组、7-羟基香豆素组（1 mmol/L）、绿原酸组（1 mmol/L）、柠檬苦素组（50 μ mol/L  ）。正常组由分离的正常小鼠子宫平滑肌细胞组成，每组重复3次。^{细胞收缩及P2X3和Ca 2+}的表达体外测定采用免疫荧光染色和激光共聚焦；采用ELISA法检测7-羟基香豆素、绿原酸、柠檬苦素给药24 h后PGE2、ET-1、NO含量。结果。代谢组学结果表明，在粗ER和PER的提取物中鉴定出7种不同的化合物，包括绿原酸、7-羟基香豆素、羟基吴茱萸碱、劳丹苷、吴茱萸碱A、柠檬苦素和1-甲基-2-[(z)-4 -壬烯基]-4(1H)-喹诺酮。体外结果显示，7-羟基香豆素、绿原酸、柠檬苦素能够抑制痛经小鼠子宫平滑肌细胞的细胞收缩和PGE2、ET-1、P2X3、Ca ²⁺，增加NO含量。结论。^{我们的研究结果表明，PER 的化合物与原始 ER 的化合物不同，7-羟基香豆素、绿原酸和柠檬苦素可以改善内分泌因子和 P2X3-Ca 2+}关闭子宫平滑肌细胞收缩的小鼠的痛经。途径。