石竹烯氧化物於酒糟鼻皮膚炎的應用評估

The Evaluations of Caryophyllene Oxide as A Potential Therapeutic Agent in Rosacea

湯澤平(TSE-PING TANG)

指導教授 ： 王家琪

高雄醫學大學/藥學院/藥學系碩士在職專班/碩士(2018年)

摘要

酒糟鼻皮膚炎主要是以面部為發病部位的一種慢性皮膚炎疾病，主要的致病原因包括先天免疫調節異常及環境因素所觸發的皮膚神經血管發炎病變。石竹烯氧化物caryophyllene oxide (CRYO)是一種富含於許多種植物的倍半萜類化合物，傳統已知可用於抗菌、抗發炎及抗腫瘤等效果。本研究的目在評估石竹烯氧化物CRYO做為治療酒糟鼻皮膚炎藥物的潛力及可能的作用機轉。
本實驗使用BALB/c小鼠活體實驗模式，先以石竹烯氧化物劑量為(1及5 mg/kg)進行小鼠腹腔注射後(一天一次) ，再以皮內注射方式施打抗菌胜肽LL-37 (320 μM, 一天兩次連續兩天)於小鼠背部皮膚的6個固定位置，觀察其誘導及治療小鼠局部酒糟鼻皮膚發炎之病灶情形。實驗於最後一次LL-37誘導後間隔12小時後取下背部病灶皮膚，其中將組織收集分別分析組織病理切片、檢測相關先天發炎因子(腫瘤壞死因子-α (TNF-α)、巨噬細胞發炎蛋白-2(MIP-2)、介白素-8 (interlukin-8))的濃度及骨髓過氧化物酶(myeloperoxidase; MPO)的蛋白質表現程度及發炎基因TNF-α及IL-8信息RNA (messenger RNA)的表現量。
活體實驗給予小鼠石竹烯氧化物CRYO藥物治療後，小鼠皮膚外觀上有減緩紅斑區範圍且局部發炎的情形，而病理組織切片中也觀察到細胞壞死及免疫細胞浸潤減少；此外，石竹烯氧化物治療後的皮膚組織分泌的腫瘤壞死因子TNF-α蛋白質表現量明顯降低，但不影響TNF-α在基因層次的表現。我們使用THP-1人類免疫單核球細胞在抗菌胜肽LL-37刺激下投予石竹烯氧化物，評估發炎細胞因子TNF-α及IL-8的蛋白質表現量。而經給予石竹烯氧化物CRYO藥物後可以抑制由抗菌肽LL-37刺激所引起的THP-1人類免疫單核球細胞所釋放的介白素IL-8造成的發炎反應。整體來說，石竹烯氧化物在酒糟性皮膚炎小鼠模型中減少了LL-37誘導的皮膚炎症反應，可能具有做為酒糟性皮膚炎治療藥物的潛力。

關鍵字

酒糟性皮膚炎 ； 石竹烯氧化物

並列摘要

Rosacea is mainly a chronic dermatitis disease with the face as the site of onset. The main causes of the disease include the abnormalities of innate immune responses and the environmental factors triggering the skin neurovascular inflammatory lesions. Caryophyllene oxide is a sesquiterpenoid that suppresses inflammation, which mainly inhibited IFN-γ productions in ovalbumin-primed splenocytes in our previous study. The purpose of this study was to evaluate the potential and possible underlying mechanisms of caryophyllene oxide as a therapeutic agent for rosacea.
The BALB/c mice were injected subcutaneously on the back with the antimicrobial peptide LL-37 (320 μM) twice daily for the production of rosacea-like symptoms for two days. Six skin lesions were generated on the back of each mouse. After the first and the third induction, caryophyllene oxide was administered at doses of 1 and 5 mg/kg by intraperitoneal injection. Twelve hours after the final induction, the skin lesions were harvested. The levels of inflammatory-related cytokines, myeloperoxidase activity and the expression of inflammatory genes were measured.
The mice treated with caryophyllene oxide significantly reduced the LL-37-induced redness and inflammatory symptoms. In histology, cell necrosis and the infiltration of immune cells were also diminished. The protein level of myeloperoxidase, tumor necrosis factor (TNF-α), macrophage inflammatory protein (MIP-2) were decreased after the treatment of caryophyllene oxide. However, caryophyllene oxide did not interfered the mRNA levels. The THP-1 human immune mononuclear cells were used to study the effects of caryophyllene oxide on human immune cells. Caryophyllene oxide can inhibit the inflammatory response in vitro by decreasing the release of interleukin-8 (IL-8) in THP-1 cell. In summary, caryophyllene oxide reduced the skin inflammation in LL-37 induced rosacea mouse model and could be a potential therapeutic agent for the treatment of rosacea.

並列關鍵字

Rosacea ； Caryophyllene Oxide

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