小葉桑根抑制豆蔻酰佛波醇乙酯誘導上皮細胞轉型之影響研究

傳統上小葉桑根被用來治療糖尿病、關節炎、風濕病，近年來被認為具有美白作用，其他生物活性還不是很清楚，天然化學預防對抗疾病作用為現代研究趨勢，我們在初步研究，證實小葉桑乙醇萃取物中具有多酚類成分，如桑辛素、氧化白藜蘆醇可能具有化學預防特性。在本研究中，我們利用 JB6 小鼠表皮細胞的模型觀察小葉桑是否有抗形成腫瘤的潛力。首先，我們先用乙醇萃取小葉桑，以試管試驗發現小葉桑根酒精萃取物具有清除 DPPH 及 NO 的能力，又在老鼠皮膚測驗中發現小葉桑根酒精萃取物具有抑制豆蔻酰佛波醇乙酯 (TPA) 誘發增生，發炎及角質化的作用，因此利用表皮細胞進行機制探討，在細胞實驗中使用 MTT 測試小葉桑根酒精萃取物及主要成分桑辛素和氧化白藜蘆醇的細胞毒性，接著選擇無毒劑量探討其對豆蔻酰佛波醇乙酯 (TPA) 去誘導 JB6 小鼠表皮細胞的細胞轉型，細胞黏附及移動的影響，結果發現小葉桑根萃取物、桑辛素和氧化白藜蘆醇可明顯去抑制 TPA 所誘導 JB6 細胞轉型，侵移及黏附。同時也會降低TPA所誘導細胞內ROS的產生、提升 GSH 的含量及抑制發炎因子 iNOS 及 COX-2 的表現，並有劑量效應。進一步在西方墨點法證實小葉桑根萃取物、桑辛素和氧化白藜蘆醇會抑制 TPA 所誘導的表皮-間質-轉化相關蛋白的表現變化，且桑辛素及氧化白藜蘆醇抑制TPA所誘導轉錄因子如 SNAIL 及 TWIST 核內含量，並且證實桑辛素及氧化白藜蘆醇可能是透過增加 GSH 含量抑制 TPA 誘導 EMT 的發生，綜合以上得知小葉桑根酒精萃取物及其主成分桑辛素及氧化白藜蘆醇可透過抗氧化活性抵抗 TPA 所誘發細胞轉型作用的能力。

關鍵字

桑辛素；促進劑；活性氧；細胞轉型

並列摘要

The roots of Morus australis have been used as a traditional Chinese medicine for the treatment of diabetes, arthritis, and rheumatism. In recent years, it evaluated as skin-whitening cosmetics. Other biological activity is not well known. In our preliminary study, it showed ethanol extract of Morus australis roots (MRE) contained polyphenolic compositions such as morusin and oxyresveratrol which may possess chemoprevention properties. Chemoprevention has been acknowledged as an important and practical strategy for the management of cancer. In the present study, whether roots of Morus australis are attributed to antitumor promotion potential are evaluated by the JB6 mouse epidermal cell model. First, ethanol extract of Morus australis roots (MRE) was prepared. MRE showed effective DPPH and NO scavenging activity in vitro. In mice skin assay, MRE inhibited 12-o-tetradecanoylphorbol-13 acetate (TPA) induced leukocyte infiltration, hyperkeratosis and hyperplasia. In JB6 cells, cytotoxicity of MRE, morusin and oxyresveratrol in JB6 cells were assessed by MTT assay. Noncytotoxic concentrations of MRE inhibited TPA induced tumorigenic anchorage-independent colonies in soft agar in a dose dependent manner. In addition, MRE, morusin and oxyresveratrol inhibited TPA-induced ROS production, epithelial-mesenchymal transitions associated protein expression and inflammatory proteins expression including iNOS and COX-2. MRE also inhibited TPA-induced actin rearrangement, cell adhesion and cell migration in JB6 cells. Furthermore, it showed that MRE, morusin and oxyresveratrol inhibited TPA-induced up-regulation of vimentin, N-cadherin and down-regulation of E-cadherin. Morusin and oxyresveratrol also inhibited TPA-induced nucler translcation of Snail and Twist which regulated epithelial-mesenchymal transitions proteins expression. In addition, the morusin and oxyresveratrol may inhibit EMT through inhibiting GSH depletion. In conclusion, our data presented that MRE has a potential of anti-transformation.

並列關鍵字

Morusin ； promoter ； reactive oxygen species ； cell transformation

參考文獻

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小葉桑根抑制豆蔻酰佛波醇乙酯誘導上皮細胞轉型之影響研究

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