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研究生:陳政憲
研究生(外文):Cheng-Hsien Chen
論文名稱:樟芝清除香菸萃取液之自由基及抑制平滑肌細胞氧化傷害
論文名稱(外文):Free radical scavenging activity and inhibiting oxidative damage effect of cigarette smoke extract on smooth muscle cell by Antrodia cinnamomea
指導教授:許世興許世興引用關係楊新玲楊新玲引用關係
指導教授(外文):Ce-shing SheuHsin-Ling Yang
學位類別:碩士
校院名稱:朝陽科技大學
系所名稱:應用化學系碩士班
學門:自然科學學門
學類:化學學類
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:159
中文關鍵詞:動脈粥狀硬化平滑肌細胞細胞凋亡自由基樟芝發酵液
外文關鍵詞:Atherosclerosissmooth muscle cellapoptosisfree radicalAntrodia cinnamomea
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第一部份
吸煙會造成人體健康的危害,主要是香煙中含有大量的自由基以及活性氧分子(ROS),導致人體內的抗氧化物質消耗而造成氧化傷害。因此若能增加體內抗氧化能力就能減少自由基或活性氧分子對人體的傷害,常見的抗氧化物例如維生素C、維生素E都是最常用來補充的抗氧化物質,本實驗中利用化學發光法(chemiluminescence)去分析香煙萃取液(CSE)中ROS的含量,並以樟芝發酵液作為抗氧化物,觀察加入樟芝發酵液之後ROS的變化;實驗結果可以看出樟芝發酵液具有清除香煙萃取液中的ROS功效,且有不錯的效果,因此樟芝在抗氧化方面也是一種很好的抗氧化物。
第二部分
吸煙容易導致粥狀動脈硬化,主要原因為LDL氧化最後形成泡沫細胞( foam cell )造成內皮損傷以及平滑肌增生,近來有研究指出平滑肌細胞凋亡也是造成粥狀動脈硬化的一個原因之一。本節實驗中採用大鼠平滑肌細胞(A7r5)探討樟芝發酵液對於加入香煙萃取液之後是否具有保護效果能夠延緩其凋亡。在comet assay實驗中發現,樟芝發酵液具有保護的效果,且香煙萃取液可能會造成SMC凋亡。western blotting的實驗中,Bcl-2加入香煙萃取液呈現減少且NF-κB也被活化,加入樟芝發酵液後Bcl-2其表現量增加且NF-κB也被抑制,Bax則變化不大。因此推測樟芝在預防平滑肌細胞的死亡具有其不錯的效果。
第三部分
吸煙一直被認為是導致的重要原因之一,已知香煙中的焦油態與氣態含有大量自由基,不但可誘發 LDL 中的多元不飽和脂肪酸產生氧化反應,也可使巨噬細胞被攝入內皮細胞中造成內皮細胞功能障礙、平滑肌增生與一連串的氧化反應,因而促進血管粥狀動脈硬化的發生。動脈粥狀硬化乃是一種慢性發炎疾病,其主要會出現在血管壁的損傷及血清中膽固醇的增加。樟芝(Antrodia cinnamomea, AC) 相傳具有強抗氧化作用的功能,且對心臟血管疾病也具有預防的功能。本實驗採用Apo E剔除(apo E knockout)公鼠,分組後餵食(cigarette smoke extract, CSE)與樟芝發酵液,並分析其總膽固醇(TC)、三酸甘油脂(TG)含量以及SOD、GPx、GRd三種抗氧化酵素活性以及總抗氧化能力TAS,並且做切片染色以觀察小鼠動脈的損傷,以及樟芝對動脈硬化是否具有保護效果。最後結果顯示出樟芝發酵液的確能夠提高TAS以及SOD、GPx、GRd的能力,並且可以降低總膽固醇、三酸甘油脂的濃度。
part1
Smoking is dangerous for human bodies.The most important reason is cigarettes contain a plenty of free radical and ROS.They induce the reduction of anti-oxidante and make damages on human bodies.All the anti-oxidante we could touch in our life is VitC and VitE;They are both used as anti-oxidante supplying human bodies.The experiment utilizes chemiluminescence to analyze the content of CSE.At first we used Fermented culture broth of A. cinnamomea (FCBA) as anti-oxidante and then observed the differences after adding FCBA.The experiment shows us that FCBA own the nice effects on clearing ROS of cigarette and it is good in thar.So we could prove the FCBA is health for us as anti-oxidante.
part2
Smoking is easy to induce artherosclerosis, because of the production of foam cells from LDL-oxidation finally make vascular endothelium damage and smooth muscle cell proliferation. Recently researches point out the cell apoptosis is the one reason of artherosclerosis.This section experiment utilized A7r5 from mats to talk about if FCBA owns the protective effects and the ability of delaying aging after adding CSE .We could observe in Comet assay FCBA indeed has protective effects,then CSE makes SMC faded.In western blotting,we find BCl-2 decreasing after adding FCBA and NF-κB been activated.On the next step we find FCBAincreasing after adding FCBA then BCl-2 been inhibiting.So we calculate that Antrodia cinnamomea is good at protecting on smooth muscel cell from dying.
part3
We all think smoking is the main reason of inducing artherosclerosis,we already knew cigarettes contain more free ions.It is not only able to activate Polyunsaturated Fat of LDL and produce oxidative-reaction,but also let macrophage be uptaked then block endothelial cell ability,smooth muscle cell proliferation and other oxidative-reaction, those responses contribute artherosclerosis. Artherosclerosis is a chronic disease.it always appears with arterial wall damage and cholesterol increase in serum.Someone tells AC owns the ability of protecting from oxidation and against cardiovascular.Our experiment utilized the atherosclerotic model of Apo E gene knock out mice,at first we grouped and feed with CSE and FCBA,then analyzed the TC,TG content,SOD,GPx,GRd anti-oxidative enzymes and TAS,finally cut into slices and dyeing in order to observe the damage of artery,and if FCBA is good at protecting from artherosclerosis.The experiment result presents FCBA indeed advance TAS and SOD,GPx,GRd ability.FCBA also could lower TC and TG.
第一部份樟芝對香煙萃取液中自由基清除的探討
摘要…………………………………………………………2-3
前言…………………………………………………………4-35
材料與方法…………………………………………………36-43
結果與討論…………………………………………………44-51
結論…………………………………………………………52-53
第二部份樟芝抑制香煙萃取液對平滑肌細胞傷害的探討
摘要…………………………………………………………54-55
前言…………………………………………………………56-75
材料與方法…………………………………………………76-91
結果與討論…………………………………………………92-106
結論…………………………………………………………107-108
第三部份樟芝對於Apo E剔除小鼠動脈硬化的探討
摘要…………………………………………………………110-111
前言…………………………………………………………112-117
材料與方法…………………………………………………118-129
結果與討論…………………………………………………130-141
結論…………………………………………………………142-143
附表…………………………………………………………144-149
參考文獻……………………………………………………150-159
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