桂皮鞣質 B1於酒糟性皮膚炎的應用評估

The Evaluations of Cinnamtannin B1 as A Therapeutic Agent in Rosacea

甘鴻霖(Hung-Lin Kan)

指導教授 ： 林英琦

高雄醫學大學/藥學院/藥學系碩士班/碩士(2017年)

摘要

酒糟性皮炎是一種以面部皮膚為主要發病部位的慢性發炎病症。先天免疫調節異常與微生物感染被認為是主要致病原因。桂皮鞣質B1 （簡稱CB1）是一種在樟樹中發現的濃縮單寧。我們先前的研究指出CB1除了本身具有抗發炎活性外，針對痤瘡桿菌與金黃色葡萄球菌也具有抗菌活性。本研究目的為評估CB1作為治療酒糟性皮炎藥物的潛力和可能機轉。活性評估採用酒糟性皮膚炎小鼠模型作測試，動物實驗在小鼠背部以皮下注射給予抗菌胜肽LL-37（320 μM），每天兩次，誘導局部的酒糟性皮膚炎病變。每隻小鼠局部產生4至6個灶症。在第一次及第三次誘導後，以腹腔注射給予劑量為20 mg / kg的CB1。在每次誘導前後，拍攝背部皮膚。在最後一次誘導後間隔20小時，取下背部灶症，以組織切片、發炎相關因子濃度測量與基因表現量評估局部發炎反應。機轉實驗則使用人類免疫單核球細胞 (THP-1)以及人類角質細胞 (HaCaT)，了解抗菌胜肽LL-37和CB1對於細胞發炎反應及活性氧化物質釋放的影響。動物實驗中給予CB1的小鼠外觀上有較小的紅斑區域並有局部發炎減少的情形，於病理切片也有較少的細胞壞死與免疫細胞浸潤。腫瘤壞死因子α（TNF-α），角質細胞趨化因子（KC）和巨噬細胞炎症蛋白-2（MIP-2）的mRNA降低，TNF-α和MIP-2的蛋白質表現量也降低。在體外試驗中，抗菌胜肽會引起人類免疫單核球細胞 (THP-1)以及人類角質細胞 (HaCaT)釋放白細胞介素 (IL-8)，而CB1可以抑制此發炎反應。此外，CB1與抗菌胜肽在抑制過氧化物誘發的活性氧化物質釋放中有增效現象。總結來說，CB1為一有潛力被作為治療酒糟性皮炎的活性成分。

關鍵字

桂皮鞣質 B1 ； 酒糟性皮膚炎

並列摘要

Background: Rosacea is a common disease characterized by chronic inflammatory of facial skin. Dysregulation of innate immunity and microbe involvement were hypothesized as key etiologies of rosacea. Cinnamtannin B1 (CB1) is a kind of condensed tannin found in Cinnamomum validinerve. Our previous study had found that CB1 have anti-inflammatory and antimicrobial activities against Propionibacterium acnes and Staphylococcus aureus. The aims of the study were to evaluate the potential of CB1 as a therapy for rosacea using a rosacea-like mice model and the potential mechanisms of the effects using in vitro cell models.
Methods: Local rosacea-like lesions were induced on the back of mice (PBS n=4; CB1 n=6) by intradermal injection of antimicrobial peptide LL-37 (320 μM) twice daily for 2 days. Each mouse had 4 or 6 lesions. CB1 at the dose of 20 mg/kg was administered intraperitoneally once daily after first LL-37-induction. The severity of the lesions was evaluated based of redness and area. At the day 3, 20 hours after the last LL-37-induction, the lesions were harvested for histology, cytokine determination, and mRNA extraction. THP-1 and HaCaT cells were used for evaluating the effects of LL-37 and CB1 on cells.
Results: The rosacea-like lesions started to be observed after the first induction. Mice treated with CB1 showed smaller redness areas and reduced local inflammation. The histology of the skin lesions indicated thickened epidermis layers and immune cell infiltrations in PBS-treated mice but not in CB1 treated mice. The mRNA of tumor necrosis factor alpha (TNF-α), keratinocyte-derived chemokine (KC) and macrophage inflammatory protein-2 (MIP-2) were decreased. Levels of TNF-α and MIP-2 in CB1-treated mice were also reduced. The in vitro tests showed that CB1 could downregulate the release of IL-8 induced by LL-37. In addition, CB1 and LL-37 showed synergistic effects on the inhibition of reactive oxygen species (ROS) induced by oxidative stress.
Conclusion: Our data indicated that CB1 have potentials to be further evaluated as a therapeutic active ingredient for rosacea.

並列關鍵字

Cinnamtannin B1 ； Rosacea

參考文獻

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