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研究生:朱智英
研究生(外文):Chih Ying
論文名稱:食茱萸粗萃物抗動脈粥狀硬化之體外研究
論文名稱(外文):In vitro experiments of anti-atherosclerosis by Zanthoxylum ailanthoides
指導教授:曾翠華曾翠華引用關係
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生化暨生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:79
相關次數:
  • 被引用被引用:3
  • 點閱點閱:573
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  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:1
隨著科技的進步生活品質也逐漸提升,由於生活習慣西化,外食機率增加,心血管疾病也進展成為十大死因之一。不少文獻指出心血管疾病的發生與過氧化作用息息相關,本實驗中以台灣本土生產的食茱萸(刺蔥;Zanthoxylum ailanthoides)作為研究,文獻指出刺蔥根莖的成分中含有抗癌、抗增生、抗病毒、抑制免疫作用、抑制血小板凝集、抗氧化、殺蟲、抗發炎、抗黴菌以及預防癌症等生物活性。然而葉部分卻無相關活性之報導。因此,本實驗室以75%酒精浸泡再以真空減壓濃縮機進行濃縮,將食茱萸粗萃物進行抗氧化成分(酚含量測定、總黃酮含量測定)、體外抗氧化活性分析(LDL electrophoretic mobility、TBARS生成濃度、apoB斷裂、共軛雙烯測定、血清膽固醇的測定、自由基捕捉測定)以及進行細胞試驗(foam cell formation)來探討食茱萸葉粗萃物與粥狀動脈硬化之間的關係。目前的實驗結果指出食茱萸葉粗萃物中含有抗氧化的多元酚類化合物(總多酚、總黃酮),食茱萸葉粗萃物可以捕捉自由基而保護低密度脂蛋白,減緩LDL的氧化,除此之外,在粥狀動脈硬化發生的過程中巨噬細胞會吞噬氧化型低密度脂蛋白進而導致泡沫細胞的形成,實驗結果指出食茱萸葉粗萃物可以減少巨噬細胞中氧化性低密度脂蛋白所造成的泡沫細胞且抑制單核球細胞轉移至血管的內膜。以上結果顯示食茱萸具有抗動脈粥狀硬化之作用。
Zanthoxylum ailanthoides is a medium to large tree, its leaves and bark are used as folk medicines to treat common cold and snakebite. It has been suggested that the stem components of Zanthoxylum ailanthoides possess bioactivities including anti-cancer, anti-poliferation, anti-viral, immuno-suppressive, antioxidant, anti-platelet, insecticidal, anti-inflamanatory, anti-fungal, anti-feedant and cancer preventative. In the present investigaion, the leaves of Zanthoxylum ailanthoides (ZLE) was extracted with 75% alcohol and condensed by vacuum pump. The anti-oxidant and anti-atherosclerosis properties of ZLE were evaluated. The results indicate that ZLE contains antioxidant components of polyphenol and flavonoid. Therefore ZLE can scavenge free radical assessed by DPPH assay. It was found ZLE can protect low lipoprotein from attacking by copper sulfate-induced free-radical and retard the oxidation of low density lipoprotein (LDL). In addition, The pathogenesis of atherosclerosis macrophage can uptake oxidative-LDL (Ox-LDL) resulting in foam cell formation. Our results indicate that ZLE can inhibit monocyte THP-1 migrating into intima and the formation of foam cell. Finally, these results suggested that ZLE decreased the CuSO4-induced lipid peroxidation and anti-atherosclerosis potential.
中文摘要..................................................1
Abstract..................................................2
壹、緒論..................................................3
一、 食茱萸之背景介紹....................................3
(一)名稱.................................................3
(二)形態特徵.............................................3
(三)分佈.................................................4
(四)成分及療效...........................................4
二、動脈粥狀硬化(atherosclerosis).......................5
三、治療與預防方向.......................................12
貳、研究動機.............................................14
參、研究架構.............................................15
肆、實驗藥品與器材.......................................16
(一)實驗試劑............................................16
(二)實驗器材、儀器......................................20
伍、實驗方法.............................................22
(一)食茱萸葉之萃取......................................22
(二)自由基捕捉測定(Determination of free radical- quenching capacity).....................................22
(三)LDL的離心萃取.......................................23
(四)Ox-LDL製備..........................................24
(五)LDL electrophoretic mobility分析....................24
(六)TBARS(MDA)生成濃度分析..............................26
(七)LDL apoB protein fragmentation assay................27
(八)酚含量測定(Total phenolic content assay) ...........28
(九)總黃酮含量測定(Total flavonoids content assay) .....29
(十)共軛雙烯測定(Diene conjugation assay) ..............30
(十一)血清膽固醇的測定(total cholesterol assay) ........30
(十二)細胞培養 (Cell Culture) ..........................32
(十三)細胞毒性分析(MTT assay) ..........................32
(十四)細胞移動性分析(Cell Migration Assay)............34
(十五)劉氏染色法(Liu,s stain)........................35
(十六)細胞分化分析(cell differentiated assay).........36
(十七)Foam cell formation assay........................37
(十八)統計分析.........................................38

陸、結果.................................................39
一、食茱萸葉之萃取與鑑定................................39
二、食茱萸粗萃取物抑制體外誘發LDL氧化之作用.............40
三、食茱萸萃取物對於Ox-LDL與單核球細胞的影響............44
四、食茱萸萃取物調控Ox-LDL對巨噬細胞的影響..............45

柒、討論.................................................49
一、食茱萸之分離與抗氧化成分分析........................49
二、食茱萸萃取物抑制LDL氧化之作用.......................49
三、食茱萸萃取物對於Ox-LDL與單核球細胞的影響............50
四、食茱萸萃取物調控Ox-LDL對巨噬細胞的影響..............51
捌、實驗結果圖表.........................................52
玖、參考文獻.............................................67
拾、參考附圖.............................................74
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