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    標題: 蓮霧花酒精萃取對於 5-Fluorouracil 抑制人類內皮細胞 EA.hy926 血管增生作用的影響
    Effects of Alcoholic Extracts from Wax Apple Flower in Combination with 5-Fluorouracil on Angiogenesis of EA.hy926 Cell
    作者: 賴貞羽
    貢獻者: 保健營養系
    楊彩秀
    劉家全
    關鍵字: 蓮霧花
    血管內皮細胞
    血管增生
    細胞移行
    細胞入侵
    基質金屬蛋白酶
    Alcoholic Extracts from Wax Apple Flower
    EA.hy926
    Angiogenesis
    Cell migration assay
    Cell invasion assay
    Matrix metalloproteinases
    日期: 2016
    上傳時間: 2016-12-21 15:36:27 (UTC+8)
    摘要: 血管增生 (angiogenesis) 是從原先已存在的微血管或位於微血管後段小靜脈所增生出血管的過程。對於大多數實質腫瘤而言,血管增生與轉移是密切相關的,藉由抑制血管增生進一步抑制腫瘤的生長,為近年來熱門探討的課題。目前5-氟尿嘧啶 (5-fluorouracil, 5-FU) 是廣泛使用在癌症上抗代謝腫瘤的治療藥物,劑量高時,可有效殺死癌細胞與轉移,但也伴隨許多副作用。因此,如何有效殺死癌細胞降低副作用,是一個很重要的課題。許多研究將癌症治療的焦點著重於血管增生的機制,並期望於日常飲食中,找出能抑制血管增生之物質,以達到平日預防的目的或合併其他癌症療法,能有良好功效。天然食品如花青素、類黃酮、 EGCG ,大部分屬於多酚類,其證實抗氧化能力得以抑制血管增生,防止正常細胞癌化等作用,降低化學治療藥物所引發的諸多副作用。達到輔助5-FU的作用,進而促進細胞凋亡的進行,使得抑制腫瘤細胞生長的效果更為顯著。本研究使用農民經疏花後丟棄的蓮霧花 (Alcoholic Extracts from Wax Apple Flower, AEWA )。目前蓮霧花研究文獻極其少數,有相關研究發現蓮霧果實抗發炎及調節血糖之功效,其功效主要來自於蓮霧果實的抗氧化活性成份。先前本實驗室證實蓮霧花具抗氧化能力及清除 DPPH 能力,且效果高於蓮霧果實本身。同時蓮霧花萃取物具有顯著美白、抗老化、抗發炎、抑制酪胺酸酶功效等功效。主要以人類臍靜脈內皮細胞 EA.hy926 與人類肺腺癌细胞株 ( A549 ) 的融合細胞,探討 AEWA 對於血管內皮細胞增生、細胞移行及入侵等相關研究,方法以細胞存活率分析 MTT assay、血管增生 In vitro angiogenesis assay、細胞移行 cell migration assay、細胞入侵 cell invasion assay、同功酵素圖譜 Zymography analysis 來測定。結果顯示 : 單獨 AEWA,( 10, 20, μg/mL ),細胞移行抑制率為13 %、46 %,入侵抑制率為12 %、12 %。AEWA ( 10, 20, μg/mL )協同5-FU,對內皮細胞的移行抑制率為43 %、76 %,入侵抑制率為43 %、67 %,皆有顯著性降低細胞移行及入侵的能力,且呈現劑量效應。結論:單獨 AEWA( 20, μg/mL ),與協同 5-FU 顯著抑制內皮細胞增生及細胞移行與入侵。AEWA可藉由抑制細胞生長及存活率,而抑制 EA.hy926 誘導之血管增生,部分與抑制血管增生分泌有關。也能輔助抗癌藥物5-FU發生協同作用,進而降低 5-FU 的用量及副作用,作為具能抑制血管增生及輔助抗癌細胞的原料,開發成保健食品。
    Angiogenesis is a kind of vessel proliferation processes from original capillary or microvascular lateral vein. For the most practical carcinoma, angiogenesis and migration have a great close relationship in each other. Recently, the most popular topics are focused on investigation of inhibiting carcinoma growth by diminishing vessel proliferation. So far, 5-fluorouracil (5-FU) is a widely used for anti-metabolic tumor in cancer treatment. 5-FU can effectively kill cancer cells and control metastasis via treatment with high doses, however, it accompanied by numerous side effects. How to effectively kill cancer cells and also reduce the side effects, become a very important issue. If the nutrients found in food can alleviate or ameliorate the side effects of anti-cancer drugs. That will be another good choice for cancer patients. Many studies of treatment for cancer emphasis on mechanism of vessel proliferation, and expect to find up the certain substance from our daily diet to decrease the vessel proliferation. The agent of the present study uses the discarded wax apple flower (WAF) from alcoholic extracts (AEWA). There is less scientific literature to examine the effects of WAF, but some researches for anti-oxidative effects of wax apple fruit are mainly concerned on anti-inflammation and regulation of blood sugar. In our previous study, the effectiveness of WAF on capacity of anti-oxidation and DPPH scavenging was better than wax apple fruit. Meanwhile, the extracts of WAF also had the significant effectiveness on whitening, anti-aging, anti-inflammation, inhibiting tyrosinase activity and so on. In this study, we attempt to investigate the effects of AEWA on proliferation, invasion and migration statuses of vascular endothelium (EA.hy926 cells, human umbilical vein endothelial cells Human umbilical vein endothelial cells (HUVEC) and human lung adenocarcinoma cells (A549) fused cells). The methods our used in this study are MTT assay, tube formation assay, cell invasion assay, cell migration assay and zymography assay and to estimate the situations of AEWA on cell viability with EA.hy926 and inhibiting vascular proliferation, invasion and migration via inhibiting and MMP- 2 activity. The findings reveal that the inhibitory rate of cell migration in single treatment of AEWA (10 and 20 μg/ml) is respectively 13% and 46%, inhibitory rate of cell invasion is 12% and 12%. Furthermore, the treatment of AEWA (10 and 20 μg/ml) accompanied with 5-FU display the 43% and 76% on the inhibitory rate of cell migration, the 43% and 67% on the inhibitory rate of cell invasion. These results indicate that no matter the single dose of AEWA or AEWA accompanied treatment with 5-FU may have significant effects on inhibiting of endothelial cell proliferation, invasion, and migration. Consequently, we speculate that AEWA probably can inhibit the EA.hy926-induced the vessel proliferation, and maybe help the effects of anti-cancer drug, 5-FU and diminish the 5-FU usages dose and side effect.
    關聯: 網際網路公開:2021-08-30,學年度:104,68頁
    Appears in Collections:[保健營養系(所) ] 博碩士論文

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