過去幾??本實驗室已指出桑椹多酚萃取物(mulberry polyphenolic extract)具有極佳的抗氧化能?，並可以抑制低密脂蛋白(LDL)的氧化；在動物實驗已經證實桑椹萃取物能抑制動脈粥?硬化(atherosclerosis)發生（附件一）、?低血脂(hyperlipidemia)與排除肝臟膽固醇堆積（附件二）；在細胞實驗?觀察到桑椹萃取物可有效抑制平?肌細胞(vascular smooth muscle cell)的增生(proliferation)與移動(migration)（附件三）且抑制高脂肪餵食之兔子之血管平?肌細胞位移及增生（附件四）。因此，我們將繼續探討桑椹多酚成份引起血管平?肌細胞增生、移動及?化(senescence)等現象而導致動脈粥?硬化的可能分子作用機轉。第一?我們將分析桑椹多酚成份並對主要多酚成份促進血管平?肌細胞NO (nitric oxide)釋放及NO造成細胞增生及位移進?研究（架構一及二）。由於Ras與血管平?肌細胞增生、位移及?化有密?的關?，第二?將重點放在桑椹多酚成份透過調控Ras?徑所引起血管平?肌細胞增生及位移及?化,包括細胞週期、移動、細胞骨架(cell cytoskeleton)及?化的分析（架構三及四）。由於Ras可能是動脈粥?硬化的重要標的並且可能受NO調控，因此第三?我們將探討桑椹多酚成份促進NO釋放是否透過抑制Ras活性而影響血管平?肌細胞增生、移動及?化，並且透過高脂肪餵食?西?兔模式得到驗證。本研究計劃?能順?達成預定目標，?僅可以?用?自桑椹的天然多酚萃取物的功能性成分達到抑制心血管病變；透過此研究?深入地瞭解動脈粥?硬化的機制並瞭解如何透過調節iNOS及Ras?預防動脈粥?硬化的發生，並評估iNOS及Ras作為治?標的可能性。 Our previous studies have shown the mulberry polyphenolic extract exerts its excellent anti-oxidative activity to inhibit LDL oxidation. Also, the mulberry polyphenolic extract has been found to inhibit atherosclerosis (supplementary Fig. 1) and reduce hyperlipidemia as well as eliminate cholesterol accumulation in experimental animals (supplementary Fig. 2). Moreover, the mulberry polyphenolic extract treatment significantly inhibits the proliferation and migration of smooth muscle cells in vitro and in vivo (supplementary Fig. 3, and 4). Therefore, we will further dissect the underling molecular mechanism of mulberry extracts involved in the proliferation, migration and senescence of smooth muscle cells that has been shown to correlate with foam cells formation, leading to atherosclerosis. The goal of the first-year of this grant project is going to identify the polyphenolic components of mulberry extract and further evaluate the effect of polyphenolic compounds on NO induction and NO-mediated the proliferation and migration of smooth muscle cells. In the second year, the function of Ras acting on inhibiting the proliferation, migration and senescence in mulberry polyphenolic extract treated cells will be verified. In the third year, we will propose the NO producing could decrease Ras activity and this phenomenon is attributable to the mulberry polyphenolic extract stimulation; meanwhile, we will establish the correlation between NO/Ras pathway and atherosclerosis. Furthermore, the efficacy of mulberry polyphenolic extract in an atherosclerotic rabbit model will be confirmed. Overall, this work will provide the mulberry polyphenolic compounds as efficient agents against cardiovascular diseases and understand the mechanism of the mulberry polyphenolic extract accounting for anti- atherosclerosis actions as well as evaluate iNOS and Ras as possible therapeutic targets.
