從流病上顯示肥胖會增加癌症發生率，尤其是肝癌，其成因之一為肥胖造成肝臟堆積脂肪，造成非酒精性脂肪肝病(NAFLD)及非酒精性脂肪肝炎病變(NASH)；另一個原因則是加強了肥胖因子如IL-6、TNF等之作用。所以降低肝臟脂肪堆積及肥胖之天然萃取物應可用來預防或治療肥胖促進肝癌之作用。來自乾燥桑葉(Morus spp.) 中的桑葉多酚萃取物(MLPE)被證實具有降血糖、降血脂、對抗粥狀動脈硬化(附件一) 等心血管疾病危險因子，我們也發現MLPE可活化AMPK促進肝癌細胞凋亡(附件二)及抑制脂肪細胞分化(附件三)，因此本計劃擬以MLPE探討抑制肥胖、NAFLD及肥胖促進肝癌作用，進一步探討是否降低肥胖性肝癌之抗藥性。本計劃工作探討桑葉多酚萃取物(MLPE)抑制(一)NAFLD及肥胖作用；(二)進行MLPE抑制肥胖促進diethylnitrosamine (DEN)致肝癌作用；(三)探討減少肥胖性肝癌之抗藥性，其架構如附圖。第一年進行(1)MLPE抑制油酸促進肝細胞脂肪堆積作用及機轉；(2)MLPE抑制脂肪細胞分化；(3)MLE及MLPE抑制高脂肪飲食誘導之非酒精性肝脂肪病變及肥胖。第二年進行(1)MLPE抑制脂肪細胞分化培養液促進肝癌細胞增生及轉移；(2)MLE抑制脂肪促進DEN致肝癌之動物試驗；(3)用蛋白質體找出促進Target及MLPE抑制機轉。第三年進行(1) 建立抗藥性肝癌細胞株R-HepG2;(2) MLPE減少脂肪細胞分化培養液促進肝癌細胞及xenograft老鼠之抗藥性之作用及機轉；(3)利用蛋白質體找出促進抗藥的Target及MLPE抑制機轉。本研究計劃若能達成預期成果，不僅可以提供桑葉抑制肥胖、NAFLD、NASH及肥胖促進肝癌之作用，且有效的助肝癌之治療。 Epidemiological studies indicated that obesity is associated with increased cancer risk. The connection between obesity and liver cancer is particularly strong. Obesity often caused by liver diseases such as non-alcoholic fatty liver disease (NAFLD) and severe non-alcoholic steatohepatitis (NASH). It is important to develop specific pharmacological or nutritional agents to reduced obesity and prevent liver disease. Mulberry leaf polyphenol extracts (MLPE) were extracted from dry mulberry leaves (Morus spp.), which have been known to modulate serum fasting glucose, lipid and atherosclerosis. (supplementary data 1). Furthermore, we also observed the effect of MLE and MLPE on stimulating hepatocarcinoma cells apoptosis and inhibiting adipocytes differention after treatment with the MLE and MLPE (supplementary data 2 and 3). According to these results, we will further study and develop the MLPE as healthy foods which were applied to attenuate NAFLD, obesity, obesity- promoted hepatocarcinogenesis, and drug resistance. The research frameworks are shown as follow figure. In first year, we will study the inhibitory effects of MLPE on (1) oleic acid- induced hepatic lipid accumulation; (2) adipocyte differentiation; (3) induction of obesity and NASH by rats fed with high-fat diet and MLE. In second years, our studies focus on (1) effects of MLPE on culture medium form differentiation adipocyte promoted proliferation and metastasis in hepatocarcinoma cells; (2) inhibition of obesity promoted diethylnitrosamine (DEN)-induced hepatocarcinogenesis in rats; (3) targeting by obesity promoted tumorgenesis. In third year, the important effects of MLPE on drug resistance caused by adipocyte differentiated culture medium in human hepatocarcinoma R-HepG2 cells and (2) xenograft mice；(3) targeting by obesity promoted doxorubicin resistance. The results of this study may demonstrate the effects of MLPE on decreasing NAFLD, NASH, and obesity. We will confirm the mechanisms of MLPE on inhibiting obesity- promoted hepatocarcinogenesis and drug resistant
