臺灣博碩士論文加值系統

經痛是一種育齡婦女常見的婦科疾病，常使用非類固醇類抗發炎藥（NSAIDs）來舒緩此症狀。此類藥物雖可經由抑制環氧酶（COX），降低前列腺素的產生，達到快速且有效的經痛舒緩效果，卻有明顯的副作用產生。因此，使用天然中草藥的替代方法，是被高度期待的。
皮薩草（oregano；Origanum vulgare L.）及香薷（herba moslae；Mosla chinensis）為兩種廣泛使用的中草藥。本研究首先測試皮薩草精油（OEO）、香薷精油（HMEO）及其主要化合物，是否具有抑制大鼠離體子宮收縮的能力，達到舒緩經痛的功效。實驗結果顯示，OEO對催產素、前列腺素F2α（PGF2α）和氯化鉀所誘發的子宮肌肉收縮分別具有29－79%（10－40 μg/ml）、8－62%（20－40 μg/ml）和45－72%（20－80 μg/ml）的抑制效果。HMEO對催產素、PGF2α和氯化鉀所誘發的子宮肌肉收縮分別有7－52%（10－40 μg/ml）、9－72%（20－80 μg/ml）和9－79%（20－80 μg/ml）的抑制效果。進一步由氣相層析質譜儀鑑定OEO及HMEO的主要化合物成分為香芹酚（carvacrol）、麝香草酚（thymol）、對異丙基甲苯（p-cymene）、沉香醇（linalool）、石竹烯（β-caryophyllene）、肉桂醛（cinnamaldehyde）及檸檬烯（limonene）。Carvacrol和thymol對催產素和PGF2α所誘發的肌肉收縮分別具有最佳的91%和87%（20 μg/ml樣品；催產素誘發）及89%和95%（80 μg/ml樣品；PGF2α誘發）的抑制效果。Thymol、linalool及carvacrol則在40 μg/ml的濃度下，對氯化鉀所誘發的肌肉收縮分別具有最佳的72%、50%和49%抑制效果。
另一方面，子宮內膜異位症和前列腺素的產生有關，進一步造成經痛的情形。為了證明上述樣品緩解經痛，也可能經由抑制子宮內膜異位症和/或降低COX-2的生成，我們以人類子宮內膜癌細胞RL95–2作為實驗模式。癌細胞分別與50 μg/ml的上述樣品作用48小時後，HMEO、cinnamaldehyde和limonene對細胞增生，分別顯示34－78%（25－100 μg/ml）、14－80%（2.5－10 μg/ml）及20－80%（10－50 μg/ml）的抑制力。除此之外，limonene尚具有明顯抑制COX–2表現量的效果。
從這些結果可以得知，OEO、HMEO、carvacrol、thymol、p-cymene、linalool、cinnamaldehyde及limonene，對子宮細胞的受體和電位調控之鈣離子通道、內膜細胞增生、COX-2表現等具有不同程度的抑制效果。因此，這些天然化合物可能對原發性及繼發性經痛的減緩及預防有良好的功效。

Dysmenorrhea, painful menstrual cramps, is the most common gynecological disorder in women of reproductive age. Non-steroid anti-inflammatory drugs (NSAIDs) interfere with prostaglandin production by inhibiting cyclooxygenase (COX) are the most commonly used therapeutic modalities for the pain. In spite of their effectiveness, the presence of side effects from these drugs limits their clinical use. A feasible alternative of natural herb remedy is therefore to be considered.
Oregano (Origanum vulgare L.) and Herba moslae (Mosla chinensis) has been widely used in traditional Chinese medicine. In this study, we firstly investigated the antispasmolytic effects of these essential oils and their main constitutes on isolated rat uterus. Essential oil of oregano (OEO) inhibited 28–79% (10－40 μg/ml), 8–62% (20–80 μg/ml) and 45–72% (20–80 μg/ml) of contractions induced by oxytocin, prostaglandin F2α (PGF2α) and KCl, respectively. Essential oil of herba moslae (HMEO) inhibited 7–52% (10–40 μg/ml), 9–79% (20–80 μg/ml) and 27–60% (20–80 μg/ml) of contractions induced by oxytocin, PGF2α and KCl, respectively. The main constituents of these essential oils were carvacrol, thymol, p-cymene, linalool, β-caryophyllene, cinnamaldehyde and limonene as identified by gas chromatography-mass spectrometry. Carvacrol and thymol showed the strongest inhibition of contractions induced by oxytocin (91% and 87%, 20 μg/ml) and PGF2α (89% and 95%, 80 μg/ml). Thymol, linalool and carvacrol showed the strongest inhibition induced by KCl (72%, 50% and 49%, 40 μg/ml).
On the other hands, dysmenorrhea associated with endometriosis is reportedly caused by increased prostaglandin production from the implants of endometriosis. In order to verify above samples relieve such dysmenorrhea by inhibiting the growth of endometriosis and/or decreasing COX-2 production, an epithelial cell line derived from human endometrium, RL95-2 cells, were used as a model. After incubation 50 µg/ml of above samples with RL95-2 cells for 48 hr, HMEO, cinnamaldehyde and limonene showed the strongest inhibitions of 34－78% (25－100 μg/ml), 14－80% (2.5－10 μg/ml) and 20－80% (10－50 μg/ml) on cells proliferation. Furthermore, limonene showed obvious inhibition on COX-2 expression.
These results suggest that OEO, HMEO, carvacrol, thymol, p-cymene, linalool, cinnamaldehyde and limonene have multiple effects on blocking the growth of endometriosis, COX-2 expression, receptor- and voltage-operated calcium channels, and then these compounds might be useful as natural agents for preventing primary and secondary dysmenorrhea.

目 錄
壹、中文摘要 1
貳、英文摘要（Abstract） 3
參、前言 5
一、皮薩草 5
二、香薷 6
三、精油 7
四、皮薩草及香薷精油的主要化學成分 7
（一）Carvacrol、thymol及p-cymene 7
（二）Linalool 8
（三）β-Caryophyllene 9
（四）Limonene 9
（五）Cinnamaldehyde 10
四、經痛類別 10
四、經痛的生理機制 12
五、原發性經痛的西藥治療 14
六、繼發性經痛的治療 14
七、子宮內膜癌 15
肆、研究動機與策略 16
伍、材料與方法 17
一、實驗材料 17
（一）精油與化學藥劑 17
（二）溶液配法 18
（三）儀器 19
（四）實驗動物 21
二、大鼠離體子宮肌肉製備 21
三、子宮肌肉收縮實驗 22
四、精油化學成分分析 22
五、子宮內膜癌細胞增生測試 23
六、西方墨漬法 23
陸、結果 25
一、皮薩草及香薷精油抑制大鼠子宮平滑肌收縮 25
二、皮薩草及香薷精油的化學成分分析 25
三、皮薩草及香薷精油的主要化學成分抑制大鼠子宮平滑肌收縮 26
四、皮薩草及香薷精油抑制人類子宮內膜癌細胞增生 27
五、皮薩草及香薷精油的主要化學成分抑制人類子宮內膜癌細胞增生 27
六、皮薩草及香薷精油的主要化學成分對人類子宮內膜癌細胞COX-2表現量的影響 28
柒、討論 29
捌、參考文獻 34
玖、表 50
拾、圖 52


表 目 錄
表一、OEO的主要化學成分 50
表二、HMEO的主要化學成分 51



圖 目 錄
圖一、皮薩草植物圖 52
圖二、香薷植物圖 53
圖三、皮薩草及香薷精油的主要化學成分結構式 54
圖四、平滑肌收縮機制 55
圖五、前列腺素生合成路徑與非類固醇類消炎止痛藥作用機制 56
圖六、催產素誘發子宮平滑肌收縮圖 57
圖七、PGF2α誘發子宮平滑肌收縮強度圖 58
圖八、氯化鉀誘發子宮平滑肌收縮圖 59
圖九、OEO及HMEO抑制催產素、PGF2α以及氯化鉀刺激物所誘發的子宮平滑肌收縮 60
圖十、OEO、HMEO及其主要成分抑制催產素誘發子宮肌肉收縮 61
圖十一、OEO、HMEO及其主要成分抑制PGF2α誘發子宮肌肉收縮 62
圖十二、OEO、HMEO及其主要成分抑制氯化鉀誘發子宮肌肉收縮 63
圖十三、OEO及HMEO抑制RL95-2細胞增生 64
圖十四、OEO、HMEO及其主要成分抑制RL95-2細胞增生 65
圖十五、OEO、HMEO及其主要成分對RL95-2細胞之COX-2表現量影響 66

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