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研究生:簡仲陽
研究生(外文):Jian, Zhong-Yang
論文名稱:樟芝萃取液對腦缺血大鼠大腦皮質脂質過氧化作用、抗氧化酵素活性與微量元素濃度的影響
論文名稱(外文):Influence Of Antrodia Camphorata (AC-10) On Lipid Peroxidation, Antioxidant Enzyme Activity, And Trace Element Levels In The Brain Cortex Of Cerebral Ischemic Rats
指導教授:林明政林明政引用關係
指導教授(外文):Lin, Ming-Cheng
口試委員:許振原方光明
口試委員(外文):Sheu, Jeun-YuanFang, Kwang-Ming
口試日期:2013-07-11
學位類別:碩士
校院名稱:中臺科技大學
系所名稱:醫學檢驗生物技術系碩士班
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:163
中文關鍵詞:腦缺血脂質過氧化作用抗氧化酵素微量元素樟芝萃取液
外文關鍵詞:cerebral ischemialipid peroxidationantioxidant enzymetrace elementAntrodia Camphorata
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在美國與台灣的老年人口中,腦中風是名列第三大死因的疾病。證據顯示:腦缺血不但會產生大量的活性氧化物與自由基,更會進一步導致細胞脂質過氧化作用。因此,大腦中適當的微量元素濃度和抗氧化酵素活性對大腦功能是非常重要的。樟芝是台灣特有的中藥材。先前的研究指出:樟芝 (AC-10) 具有多種生物活性作用,包括抗發炎,抗氧化和抗腫瘤。到目前為止,AC-10 萃取液對於腦缺血時微量元素濃度、脂質過氧化狀態和抗氧化酵素活性的影響尚未被研究,因此,需要更進一步予以探討。而透過我們實驗結果顯示:於腦缺血之前給予 AC-10不僅可以顯著降低大鼠大腦皮質內 MDA 濃度與微量元素包括鐵 (Fe)、銅 (Cu)、鉛 (Pb) 和鉻 (Cr) 的濃度。相反地,血漿內抗氧化酵素活性包括:超氧化物歧化酶 (SOD)、觸酶 (CAT)、麩胱甘肽過氧化酶 (GPX) 與大腦皮質鋅 (Zn) 濃度則顯著上升。總而言之,我們的實驗結果證明:AC-10不但能在腦缺血情況下降低脂質過氧化產物之 MDA 濃度,同時也能減少微量元素鐵 (Fe)、銅 (Cu)、鉛 (Pb) 和鉻 (Cr) 濃度而發揮其保護作用。此外,AC-10在腦缺血組手術前的預防組大鼠中可增強抗氧化酵素活性以及鋅 (Zn) 濃度,最重要的是這種正面效應被認為是有利於改善在腦缺血後所引起的氧化性傷害。
Cerebral ischemia is the third leading cause of death in older population in the United States and Taiwan. Evidence revealed that cerebral ischemia not only may generate a large amount of reactive oxygen species but also may eventually lead to further oxidative lesion to the brain. Proper trace element level and antioxidant enzyme activity is recognized to be important to the brain function. Antrodia Camphorata (AC-10) is one of the unique Chinese medicines especially found in Taiwan. Previous study indicated that AC-10 possesses a variety of biological effects including anti-inflammatory, anti-oxidation and anti-cancer. So far, biological effects of the extract of AC-10 on the alteration of trace element level, lipid peroxidation status, and antioxidant enzyme activity in the ischemic brain, however, has not been investigated and needs to be further elucidated. Our experimental results showed that pretreatment rats with AC-10 prior to cerebral ischemia not only may significantly decrease lipid peroxidation intensity but also may decline the trace element level including iron (Fe), copper (Cu), lead (Pb), and chromium (Cr) as compared with the ischemic rats. In contrast, antioxidant enzyme activity of superoxide dismutase, catalase, glutathione peroxidase in the plasma and trace element level of zinc (Zn) in the homogenates of brain cortex of rats were significantly increased in AC-10 pretreatment rats as compared with the ischemic rats. Taking all evidence together, a possible mechanism we manifest here that AC-10 seems to exert its beneficial effects not only in declining lipid peroxidation status but also in reducing trace element level of Fe, Cu, Pb, and Cr in the ischemic brain. Moreover, pretreatment rats with AC-10 prior to cerebral ischemic surgery may enhanced antioxidant enzyme activity and Znic (Zn) level in the ischemic brain and crucially, this positive effect is also believed to be helpful for the brain in ameliorating oxidative injury caused by cerebral ischemic event.
中文摘要 1
英文摘要 2
第一章 前言
第一節 研究動機與策略
第二章 文獻回顧
第一節 牛樟芝
一、 樟芝由來與命名 7
二、 樟芝分布區域與形態特徵 8
三、 樟芝與其主要活性成分 8
1、 樟芝多醣體 10
(1)、 多醣體種類分析 10
(2)、 多醣體活性分析 11
2、 樟芝三帖類 12
(1)、 三萜類生成機制 12
3、 樟芝抗氧化物質 13
四、 樟芝藥理活性之應用 13
1、 護肝功能 13
2、 提升免疫功能 14
3、 抗氧化能力 14
4、 抑制腫瘤細胞生長 15
5、 藥理毒性試驗 15
第二節 自由基與活性氧定義
一、 自由基 (Free radical) 16
二、 活性氧 (Reative oxygen species) 17
三、 自由基種類 18
1、 超氧化物 (Superoxide, O2•-) 18
2、 氫氧自由基 (Hydroxy redical, OH•) 19
3、 過氧化物自由基 (Peroxy radical, ROOH•) 19
4、 過氧化氫 (H2O2) 19
5、 一氧化氮 (NO•) 19
6、 過氧亞硝基陰離子 (ONOO-) 20
四、 脂質過氧化作用 (Lipid peroxidation) 20
1、 起始反應期 (Initiation of autoxidation) 21
2、 連鎖增殖反應期 (Propagation of autoxidation) 21
3、 終止反應期 (Termination of autoxidation) 22
五、 脂質過氧化與疾病 22
第三節 氧化性傷害 (Oxidative damage) 之相關性探討
一、 活性氧與自由基對細胞之氧化傷害概述 24
1、 對脂質之傷害 24
2、 對蛋白質之傷害 25
3、 對DNA之傷害 26
二、 氧化性傷害與疾病之相關性 27
三、 氧化性傷害與老化之相關性 27
四、 氧化性傷害對於生物體之影響 28
第四節 抗氧化防禦系統種類介紹
一、 非酵素性防禦系統之抗氧化物與抗氧化劑 29
1、 維生素C (Ascorbic acid) 30
2、 維生素E (Vitamin E) 31
3、 麩胱甘肽 (Glutathione, GSH) 32
4、 β-胡蘿蔔素 (β-carotene) 32
5、 類黃酮 (Flavonoids) 33
二、 抗氧化劑種類與作用機制 34
1、 自由基終止劑 (Free radical terminators) 34
2、 還原劑或氧的捕捉劑 (Reducing agent or oxygen scavenger 34
3、 金屬螯合劑 (Metal chelator) 35
4、 單旋態氧之消光劑 (Singlet oxygen quencher) 35
5、 相乘劑 (Synerigist) 36
三、 酵素性防禦系統之抗氧化物 37
1、 超氧化岐化物 (Superoxide dismutase, SOD) 37
2、 麩胱甘肽過氧化化酶 (Glutathione peroxidase, GPX) 38
3、 觸酶 (Catalase, CAT) 39
4、 麩胱甘肽還原酶 (Glutathione reductase, GRD) 40
5、 麩胱甘肽硫轉移酶 (Glutathione S-transferase, GST ) 40
第五節 微量元素
一、 微量元素之種類 42
二、 微量元素各論探討 44
1、 鋅 (Zinc, Zn) 44
2、 鐵 (Iron, Fe) 45
3、 銅 (Copper, Cu) 45
4、 硒 (Selenium, Se) 46
三、 重金屬元素 47
1、 鉛 (Lead, Pb) 47
2、 鉻 (Chromium, Cr) 48
第三章 材料與方法
第一節 實驗材料
一、 實驗動物 50
二、 樟芝萃取液製備 50
三、 樟芝萃取液 (AC-10) 51
四、 化學試藥 52
五、 分析檢驗套組 53
六、 儀器設備 53
第二節 實驗設計與流程
一、 實驗設計 56
二、 實驗分組 56
三、 大鼠腦中風手術 57
四、 血液、臟器之收集 57
五、 各臟器組織液處理 58
第三節 分析方法
一、 超氧化物歧化酶 (Superoxide dimutase, SOD) 酵素活性測定 59
二、 觸酶 (Catalase, CAT) 酵素活性測定 61
三、 麩胱苷肽過氧化酶 (Glutathione peroxidase, GPX) 酵素活性測定 63
四、 丙二醛 (Malondialdehyde, MDA) 濃度之測定 65
五、 蛋白質 (Protein) 濃度之測定 66
六、 石墨爐式原子吸收光譜儀 (GFAAS) 分析測定 67
七、 統計學分析 71
第四章 結果
第一節 血漿內抗氧化酵素活性測定結果
一、 大鼠血漿之超氧化物歧化酶 (Superoxide dimutase, SOD) 活性測定結果 72
二、 大鼠血漿之觸酶 (Catalase, CAT) 活性測定結果 72
三、 大鼠血漿之麩胱苷肽過氧化酶 (Glutathione peroxidase, GPX) 活性測定結果 73
第二節 組織中脂質過氧化產物 MDA 分析結果
一、 大鼠大腦皮質脂質過氧化產物 MDA 濃度分析結果 74
二、 大鼠肺臟之脂質過氧化產物 MDA 濃度分析結果 74
三、 大鼠肝臟之脂質過氧化產物 MDA 濃度分析結果 75
四、 大鼠腎臟之脂質過氧化產物 MDA 濃度分析結果 75
五、 大鼠睪丸之脂質過氧化產物 MDA 濃度分析結果 76
六、 大鼠血漿之脂質過氧化產物 MDA 濃度分析結果 76
第三節 大鼠大腦皮質內微量元素測定結果
一、 大鼠大腦皮質內微量元素銅 (Cu) 之分析結果 77
二、 大鼠大腦皮質內微量元素鐵 (Fe) 之分析結果 77
三、 大鼠大腦皮質內微量元素鋅 (Zn) 之分析結果 78
四、 大鼠大腦皮質內微量元素鉛 (Pb) 之分析結果 78
五、 大鼠大腦皮質內微量元素鉻 (Cr) 之分析結果 79
六、 偵測極限 (Instrument detection limit, IDL)、精密度 (Precision) 與準確度 (Accuracy) 分析結果 80
第五章 討論
一、 樟芝萃取液對腦缺血大鼠血漿內抗氧化酵素活性之效應探討 82
二、 樟芝萃取液對腦缺血大鼠中組織脂質過氧化作用效應 84
三、 樟芝萃取液對腦缺血大鼠大腦皮質內微量元素之效應 85
參考文獻 90
表 107
圖 123
附錄. 縮寫字對照表 150

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