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研究生:黃安中
研究生(外文):An—Chung Huang
論文名稱:洛神花花青素萃取物抗氧化型低密度脂蛋白所引發動脈粥狀硬化的機轉
論文名稱(外文):Hibiscus Anthocyanins inhibits oxLDL-mediated macrophage apoptosis and CD36 expression.
指導教授:王朝鐘王朝鐘引用關係
指導教授(外文):Chau - Jong Wang
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:91
中文關鍵詞:低密度脂蛋白動脈粥狀硬化
外文關鍵詞:low density lipoproteinatherosclerosis
相關次數:
  • 被引用被引用:10
  • 點閱點閱:334
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:1
近年來,科學家在追尋抗氧化、抗衰老的過程中,發現在一些高等植物中所含之多酚化合物,具有抗氧化作用,藉由抗氧化的作用來抑制LDL的氧化修飾,是一種有效預防動脈粥狀硬化發生的方法。本研究之目的為評估由洛神花所分離的花青素萃取物: HAs,其抗LDL氧化及抗動脈粥狀硬化之作用效果,而進行體外抗氧化實驗與細胞實驗來驗證。
本實驗以乾燥洛神花為材料,以調酸的甲醇溶劑萃取其花青素,發現甲醇萃取率為5%;純度測定約有85~95%,另以HPLC分析後發現其花青素主要分為兩類,推測分別為delphinidin系及cyanidin系,其花青素含量分別為3.7%及0.9%。
以銅離子誘導LDL氧化的模式進行體外抗氧化實驗,並以LDL氧化時其蛋白部分的氧化修飾反應與脂質部分的過氧化反應作為氧化指標。藉由ApoB衍蛋白斷裂現象及LDL蛋白表面電荷改變的程度與脂質過氧化產物MDA的生成量減少的結果發現,花青素萃取物具有抑制LDL氧化的能力,此外也具有清除1,1-diphenyl-2-picrylhydrazyl(DPPH)自由基的作用。而在細胞實驗方面,花青素萃取物具有保護巨噬細胞不受因LDL氧化作用而死亡;而在給予低濃度的花青素萃取物時,會使OxLDL所引起的CD36表現有下降的情形,進而減少巨噬細胞轉形為泡沫細胞(foam cell),這些現象都會減緩動脈硬化的發生過程。
由上述體外抗氧化實驗與細胞實驗結果顯示,花青素萃取物具有極佳的抗氧化活性並能抑制LDL的氧化,此外也參與調控巨噬細胞因LDL的氧化而產生促進動脈粥狀硬化生成的反應。綜合以上結果,我們認為花青素萃取物具有預防動脈粥狀硬化的作用,而能應用於保健食品的開發。
Recent investigations highlight the antioxidant components of plant that may decreased the risk of inflammatory diseases and certain forms of cancer.The oxidative modification of low density lipoprotein plays a key role in the pathogenesis of atherosclerosis. Antioxidative reagents, which can effectively inhibit LDL oxidation, may prevent atherosclerosis via reducing early atherogenesis, and slowing down the progression to advance stages.
The major material of this experiment is anthocyanins which was extracted from dried Hibiscus sabdariffa L. by the acidic methanol. We found that only 5% anthocyanins could be extracted by acidic methanol as well as the purity could reach 85 to 90%. Two categories of anthocyanins were identified by the HPLC analysis. One is delphinidin that takes 3.7% of anthocyanins extraction, and the other one is cyanidin which holds 0.9% of anthocyanins extraction.
As shown in previous studies Hibiscus sabdariffa L. is a nature plant containing a lot of pigments that was found to possess antioxidative activity.In this study, we evaluated the antioxidative activity of the Hibiscus anthocyanins (HAs) by measuring their effects on LDL oxidation (in vitro) and anti-atherosclerotic abilities. The antioxidative activity of the extracts on LDL oxidation was defined by Apo B fragmentation, relative electrophoretic mobility (REM) and thiobarbituric acid-relative substances (TBARS) assay. Our results showed that HAs were able to inhibit the Apo B fragmentation, REM and TBARS assay in the Cu2+-mediated oxidative LDL. HAs also possessed the ability of DPPH radical scavenging. Taken together, HAs showed strong potency to inhibit the LDL oxidation induced by copper. When given low concentration of HAs, the expression of CD36 induced by OxLDL was reduced. Further studies demonstrated that HAs could reduce the cholesterol accumulation in macrophage, and that,subsequently, inhibit the formation of foam cells.
In conclusion, Hibiscus anthocyanins (HAs) possess strong antioxidative ability to inhibit LDL oxidation and foam cell formation. Therefore, it is suggestive the anthocyanins could be a healthy food to prevent individuals from atherosclerosis.
壹、中文摘要...............................................2
貳、英文摘要...............................................4
參、緒論...................................................5
一、洛神花之背景介紹.......................................5
二、花青素之背景介紹.......................................7
三、動脈粥狀硬化(atherosclerosis)........................8
肆、研究動機..............................................15
伍、研究架構..............................................17
陸、實驗方法..............................................18
柒、實驗藥品與器材........................................31
捌、結果..................................................35
一、洛神花花青素之分離與鑑定..............................35
二、洛神花花青素萃取物抑制體外誘發LDL氧化之作用...........36
三、洛神花花青素萃取物調控oxLDL對巨噬細胞的作用影響.......39
四、洛神花花青素萃取物調控oxLDL所引發動脈粥狀硬化的機轉...42
玖、討論..................................................45
一、洛神花花青素之分離....................................45
二、洛神花花青素萃取物抑制LDL氧化之作用...................46
三、洛神花花青素萃取物調控oxLDL對巨噬細胞的作用影響.......47
四、洛神花花青素萃取物調控oxLDL所引發動脈粥狀硬化的機轉...48
拾、實驗結果圖表..........................................51
拾壹、參考附圖............................................71
拾貳、參考文獻............................................80
拾参、附錄................................................88
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