一、具促進皮膚傷口癒合之植物萃取物回顧 二、蘭花萃取物對皮膚傷口癒合效用之探討

1. Review of botanic extracts with wound healing function 2. Wound healing efficacy of orchid extract

張舒涵(Shu-Han Chang)

指導教授 ： 翁家瑞

台南應用科技大學/生活科技學院/生活服務產業系生活應用科學碩士班/碩士(2018年)

摘要

傷口的定義為皮膚受到外來性的刺激、破壞或損傷。傷口癒合是一種複雜的生理反應過程，當皮膚組織受到傷害時，皮膚會啟動癒合機制，過程包含發炎、增生及重塑。目前有許多植物萃取物含有抗發炎、抗氧化等功能，在化妝品及皮膚保健產品上被廣大應用。當皮膚受到創傷時，傷口癒合過程可分為發炎反應期、組織增生期及組織重塑期；其中增生期可利用基質金屬蛋白酶 (matrix metalloproteinase, MMP) 清除細胞外基質，重塑期則可透過轉化生長因子 (transforming growth factor beta 1, TGF-β1) 分泌誘發第一型膠原蛋白 (type I collagen) 產生。若能刺激 TGF-β1 分泌，促使皮膚成纖維細胞產生細胞移行並促進膠原蛋白生成，可加速皮膚傷口癒合。靈芝醣肽多醣體 (polysaccharide) 被證實可促進蛋白質生成，利於傷口癒合；而蘭科植物中亦被發現含有醣肽多醣體，在先前的研究中發現有抗發炎的效果，但其對傷口癒合效用及作用機制的研究目前仍有限。本研究分為兩部分，第一部分為歸納已被證實具有皮膚傷口癒合功效的植物萃取物，並整合這些可促進皮膚傷口癒合的科學根據；第二部分為體外細胞試驗，探討蘭花萃取物 (water extract of Phalaenopsis orchid flower, WEPF) 對皮膚傷口癒合相關作用機制。本研究方法以 MTT assay 測定 WEPF 對細胞之毒性，Western blotting 測定蛋白表現量，wound healing assay 檢測細胞移行情形。結果顯示，濃度 200 μg/mL 以下之 WEPF 對皮膚成纖維細胞 CCD-966SK 處理 48 小時，並無顯著毒殺作用；經 WEPF 處理細胞 48 小時，發現細胞有顯著移行作用；另外，WEPF 可能透過活化 TGF-β/SMAD 訊息路徑誘發 type I collagen表達並抑制 MMP-1 蛋白。綜合以上，WEPF 可抑制 CCD-966SK 細胞 MMP-1 蛋白量，保留type I collagen 含量，並可促使細胞移行，達成促進皮膚傷口癒合的效用。

關鍵字

蘭花萃取物 ； 傷口癒合 ； 細胞移行 ； 轉化生長因子 ； 第一型膠原蛋白

並列摘要

The wound is defined as exogenous irritation, destruction or damage to the skin. Wound healing is a complex physiological reaction that initiates a healing mechanism when the skin tissue is damaged. The process involves inflammation, hyperplasia, and remodeling. At present, many plant extracts with anti-inflammatory and anti-oxidative activities are widely used in the cosmetics and skin care products. When skin is injuried, wound healing will initiate and the processes can be divided into the inflammatory stage, the proliferative stage, and the remodeling stage. In proliferative stage, matrix metalloproteinase (MMP) is used to clear the extracellular matrix. In remodeling stage, type I collagen is induced by the TGF-β1 (transforming growth factor beta 1). The wound healing of skin should be accelerated if the TGF-β1 is stimulated to promote skin cell migration and collagen production. Polysaccharide was proved that benefited to wound healing by promoting protein production. Orchid plants were found that containing glycopeptide polysaccharides and having anti-inflammatory effect, but the mechanism of action is still unclear. There are two parts in this study. In the first part, the related literatures concrerning natural products with wound healing function were collected and the data were analyzed and concluded. In the second part, a water extract of Phalaenopsis aphrodite subsp. Formosana (WEPF) was used to evaluate the healing effect and mechanism on CCD-966SK skin fibroblasts. The toxicity of WEPF on CCD-966SK cells was assayed by MTT method. The levels of protein were analyzed by Western blots. The cell migration was detected by wound healing assay. The results showed that WEPF at concentrations less than 200 μg/mL was untoxic to CCD-966SK cells in 48 hrs treatment. By treating 100 μg/mL of WEPF on CCD-966SK cells for 48 hrs, a remarkable migration was observed. Moreover, the protein levels of type I collagen and MMP-1 were increase and decrease, respectively, by WEPF via TGF-β/Smad pathway. In conclusion, WEPF can inhibit the MMP-1 protein and retain the type I collagen content in CCD-966SK cells as well as can promote cell migration to promote skin wound healing.

並列關鍵字

WEPF ； wound healing ； cell migration ； transforming growth factor ； type I collagen

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