山苦瓜萃取物暨其區分物對於痤瘡桿菌誘導發炎反應的影響

Effects of wild bitter melon extracts on Propionibacterium acnes-induced inflammation

李宥苡(You-Yi Li)

指導教授 ： 蔡帛蓉

國立臺灣師範大學/教育學院/人類發展與家庭學系/碩士(2012年)

摘要

痤瘡(acne)俗稱青春痘，其致病因子複雜。過度皮脂生成(sebaceous hyperproduction)、毛囊過度角質化(follicular hyperkeratinization)、痤瘡桿菌增生(Propionibacterium acnes colonization)和發炎反應(periglandular dermal inflammation)是重要的致病因子。其中痤瘡桿菌(P. acnes)被認為在痤瘡發展過程中扮演重要的角色，P. acnes藉由活化單核球細胞(monocytes)與角質細胞(keratinocytes)的toll-like receptor，啟動下游的訊息傳遞路徑產生發炎介質誘導痤瘡發炎情形；另外P. acnes誘發產生活性氧分子(reactive oxygen species, ROS) 與基質金屬蛋白酶(matrix metalloproteinases, MMP)亦參與發炎反應。苦瓜(Momordica charantia)萃取物具有抗發炎、抗氧化能力。因此，本研究欲探討台灣山苦瓜(Momordica charantia Linn. var. abbreviata Ser.; wild bitter melon)萃取物是否具有抑制P. acnes生長與其誘發之發炎反應的效用。
製備數種山苦瓜萃取物，收集山苦瓜樣品(育種之花蓮1~4號，67-11品系與野生品種)之不同部位(果實、葉、藤)，果實以溶劑(乙醇、正己烷、乙酸乙酯和水)進行萃取，而葉以甲醇進行萃取。以broth dilution method實驗的結果發現67-11與野生品種葉、藤甲醇萃物能抑制痤瘡桿菌生長。以P. acnes刺激THP-1 monocytes的模式發現山苦瓜果實乙酸乙酯和乙醇萃取物與葉子甲醇萃物具in vitro抗發炎作用。為進一步探討山苦瓜果實萃取物之區分物(fractions)的抗發炎作用，取得皂化物、非皂化物、oleic acid, linoleic acid, α-linolenic acid, conjugated-linolenic acid, phytol和lutein，結果顯示不皂化物和lutein抑制cytokine和MMP-9表現量，皆具有in vitro抗發炎作用，而長鏈脂肪酸(oleic acid, linoleic acid, α-linolenic acid, CLN)僅降低IL-1β濃度。另發現中鏈脂肪酸(capric acid和lauric acid)能夠減少P. acnes刺激THP-1 monocytes的cytokines產生。在in vivo抗發炎實驗，將P. acnes注射至ICR小鼠耳朵測試樣品的抗發炎能力，結果顯示山苦瓜果實EA萃物、capric acid、山苦瓜葉子甲醇萃物與總多酚萃取物能夠抑制P. acnes誘發的發炎反應與耳朵腫脹情形。
綜合論述，山苦瓜果實EA萃物與葉子甲醇萃物在in vitro and in vivo實驗中顯示具有抑制P. acnes誘發的促發炎介質與小鼠耳朵腫脹情形，而果實EA萃物中含的lutein與葉子中含的總多酚可能是抗發炎的有效成份。

關鍵字

痤瘡 ； 痤瘡桿菌 ； 山苦瓜 ； 發炎

並列摘要

Acne vulgaris is a common skin disease involving pilosebaceous follicle. The pathogenesis of acne vulgaris is multifactorial, including increased sebum production, comedogenesis and Propionibacterium acnes proliferation. P. acnes plays an important role not only in the process of inflammation but in the formation of comedones. Previous studies have shown P. acnes would activate monocytes to secrete pro-inflammatory cytokines, some studies have found that matrix metalloproteinases (MMPs) will participate in the progression of acne. Wild bitter melon (Momordica charantia Linn. var. abbreviata Ser.) possesses numerous pharmacological actions such as antibacterial, antioxidant, anti-diabetic, and anti-inflammatory activities. The aim of this study was to evaluate the anti-microbial activity as well as in vitro and in vivo inhibitory effect of wild bitter melons extracts and carpic acid on P. acnes-induced inflammation.
Our results showed that leaves and vine extracts from #67-11 and wild varirties of wild bitter melons effectively inhibited the growth of P. acnes. The ethyl acetate and ethanol extract from WBM fruit significantly reduced IL-8, TNF-α, IL-1β and MMP-9 levels by P. acnes-stimulated THP-1 cells. We further evaluated the fractions and composition of WBM fruit EA extract, the results showed that saponification and unsaponification fractions of WBM fruit, phytol, lutein, capric acid, and lauric acid can reduce cytokine levels. Unsaponification fraction of WBM fruit, lutein, and lauric acid can reduce MMP-9 protein levels. Fatty acids including oleic acid, linoleic acid, α-linolenic acid and conjugated linolenic acid reduced IL-1β production. In addition, ethyl acetate extract of WBM fruit, capric acid, methanolic extract and total phenol extract of WBM leaves reduced P. acnes-induced inflammation of mice ear and then showed in vivo anti-inflammatory activity.
Our results suggested that wild bitter melon extracts have anti-inflammatory effects against P. acnes and may be useful in the adjuvant treatment of acne.

並列關鍵字

acne ； Propionibacterium acne ； Momordica Charantia ； inflammation

參考文獻

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