| 摘要: | 大豆分離蛋白為目前應用於保健食品主要成分或添加物的新興來源，因其富含的胺基酸成份能有效補充身體所需的能量及強化生理代謝而受到注目。但針對其對於生理功能的改善效果仍未有充足的相關研究驗證，故本試驗利用小鼠大腸癌細胞株及大腸癌小鼠模式動物，評估經由全植物來源精萃之大豆分離蛋白，對於大腸直腸癌的改善效果與其作用機轉，並同時藉由極致效能液相層析串聯式質譜儀（Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry, UPLC Q-TOF MS/MS）及其分析軟體Progenesis QI進行代謝體分析，深入探討對於大腸癌細胞株及活體動物中代謝產物的影響。透過極致效能液相層析串聯式質譜儀及感應耦合電漿質譜儀（Inductively coupled plasma-mass spectrometer, ICP-MS），我們證明經精萃後大豆蛋白所含之主要胺基酸如arginine、leucine及lysine等純度皆高於95%，且含有磷、鈉、鈣、銅及硒元素。分別以細胞及模式動物進行細胞存活率分析、免疫組織化學切片、西方墨點法及即時聚合酶連鎖反應等實驗。結果皆顯示大豆分離蛋白確實對於大腸直腸癌具顯著抑制效果，且根據代謝體分析結果發現，大豆分離蛋白能透過調控參與mTOR代謝路徑的蛋白質（Ang-2、PLP1與AKT）達到抑制作用。
Soybean isolated protein, an emerging source, currently used as the main ingredients or additives of healthy food. Its rich in amino acids, which are able to supply body energy required and strengthen physiological metabolism effectively. However, there is still not enough relevant research to verify the improvement effect of soybean isolated protein on physiological functions. Therefore, this study evaluates the improvement effect of soybean isolated protein extracted from whole plant sources on colorectal cancer by cell lines and colorectal cancer mouse model. In addition, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC Q-TOF MS/MS) and its software Progenesis QI were used for metabolite analysis. Through the performance of inductively coupled plasmamass spectrometer (ICP-MS) and UPLC Q-TOF MS/MS, we have proved that the purities of the main amino acids, including arginine, leucine and lysine, contained in the soybean isolated protein after extraction are all over 95%. In addition, phosphorum, sodium, calcium, copper and selenium are contained as well. Cells and model animals were used for cell viability analysis (MTT assay), immune-histochemical sectioning, western blotting, and RT-PCR. Our results showed that soybean isolated protein did have a significant inhibitory effect on colorectal cancer. According to metabolite analysis results, soybean isolated protein can inhibit colorectal cancer by regulating proteins involved in mTOR metabolic pathway, such as Ang-2, PLP1 and AKT. |
