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研究生:李蒔華
研究生(外文):Shih-Hua
論文名稱:大蒜精油有機含硫成分對於內毒素誘發之大鼠小腸嗜中性球浸潤之影響及相關機制探討
論文名稱(外文):Effect of garlic organosulfur compounds on LPS-induced neutrophil infiltration in the intestine of rats and the related mechanism
指導教授:劉承慈
指導教授(外文):Cheng-Tzu Liu
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:營養學研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:79
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已知大蒜具有抗發炎作用,我們過去的研究發現大蒜精油 (GO) 可降低LPS誘發之小腸嗜中性球浸潤,這與GO降低周邊血嗜中性球密度及降低嗜中性球與血管壁黏附因子表現有關。本研究之目的為探討GO中何種有機含硫化合物為改善嗜中性球浸潤腸道組織的主要成分及其機制。實驗設計係分別以高、低兩種劑量(0.125或0.025 mmol/kg BW)GO有機含硫化合物-二烯丙基硫化物 (DAS)、二烯丙基二硫化物 (DADS)及二烯丙基三硫化物 (DATS) 給予大鼠隔日胃灌,共八次,再由腹腔注射LPS (5 mg/kg BW) 誘發系統性發炎反應 (systemic inflammation response syndrome)。注射LPS 18小時後犧牲大鼠,收集下腔靜脈血分離嗜中性球,以流式細胞儀分析細胞表面CD11b和CD18的表現;收集注射LPS前後血液 (0 hr、18 hr) ,以ELISA分析血清中可溶性黏附因子sL-selectin(souble L-selectin)、sICAM-1(souble intercellur adhesion molecule-1)和趨化因子CINC-1(cytokine-induced neutrophil chemoattractant)的含量;收集迴腸組織分析黏附因子L-selectin、ICAM-1、趨化因子CINC-1和促發炎激素TNF-α、IL-1β的含量。嗜中性球浸潤小腸組織的程度則是藉由分析迴腸組織中MPO的活性和以迴腸切片之MPO化學染色法來觀察。另外,以HE形態學染色來觀察迴腸組織的完整性。
結果顯示,在不影響大鼠正常代謝下,灌食DAS、DADS及DATS,不論高低劑量皆可改善LPS誘發之嗜中性球浸潤腸道組織的程度。DAS不論在高或低劑量,均可顯著降低周邊血嗜中性球密度、CD11b/CD18的表現、血清中sICAM-1和CINC-1的含量以及迴腸黏膜上皮組織ICAM-1、CINC-1、TNF-α及IL-1β含量,但對於降低血清中sL-selectin含量僅在低劑量時具有顯著性。DADS不論在高或低劑量,均可顯著降低嗜中性球表面CD11b表現、迴腸黏膜上皮組織ICAM-1、TNF-α及完整迴腸組織的CINC-1和TNF-α含量,但對於降低血清中嗜中性球表面CD18表現、迴腸黏膜上皮組織CINC-1、完整小腸組織ICAM-1、IL-1β含量之作用,僅在高劑量下有顯著影響。DATS不論低或高劑量下均顯著抑制嗜中性球表面CD11b表現,且在低劑量時也能顯著降低迴腸黏膜上皮組織ICAM-1、TNF-α及IL-1β含量,但在高劑量DATS下,則會惡化LPS誘發下增加之小腸CINC-1含量,並伴隨惡化之小腸受損型態。由以上結果可推論,GO含硫化物在不影響大鼠正常代謝下,均能改善LPS誘發系統性發炎反應之大鼠嗜中性球浸潤組織的程度,且在適當劑量下這些GO含硫成分可有效改善嗜中性球與血管壁黏附因子及趨化因子的表現降低促發炎細胞激素在組織中的含量,此活性提供其抗發炎作用的另一種角色。


圖表次………………………………………………………………………………I
中英文專有名詞及縮寫對照表……………………………………………………III
摘要…………………………………………………………………………………V
前言…………………………………………………………………………………VII
第一章、文獻探討
壹、系統性發炎反應 (Systemic inflammatory response syndrome)……………1
一、系統性發炎反應概論……………………………………………………………1
二、系統性發炎反應和敗血症………………………………………………………2
三、 LPS(lipopolysaccharide)與系統性發炎反應………………………………..3
貳、嗜中性球(Neutrophil)在系統性發炎反應的角色……………………………7
一、嗜中性球…………………………………………………………………………7
二、嗜中性球生命週期………………………………………………………………8
三、嗜中性球的遷移活性……………………………………………………………8
四、嗜中性之活化與組織損傷………………………………………………………15
參 、腸道免疫與嗜中性球 …………………………………………………………18
一、腸道免疫系統……………………………………………………………………18
二、嗜中性球之組織浸潤……………………………………………………………20
肆 、大蒜與其有機硫化物 …………………………………………………………21(1)大蒜在抗癌方面研究 …………………………………………………………23
(2)大蒜在抗氧化方面研究 ………………………………………………………23
(3)大蒜在抗發炎方面研究 ………………………………………………………24
(4) 大蒜對嗜中性球浸潤組織的相關研究………………………………………24
伍、研究動機與目的…………………………………………………………………26
第二章、實驗材料與方法……………………………………………………………27
壹、實驗材料與儀器………………………………………………………………27
一、化學試藥………………………………………………………………………27
二、分析套組………………………………………………………………………29
三、儀器設備………………………………………………………………………29
四、大蒜精油有機硫成分試藥……………………………………………………30
五、實驗動物………………………………………………………………………30
貳、實驗方法………………………………………………………………………31
一、實驗流程………………………………………………………………………31
二、實驗動物之處理………………………………………………………………32
1. 系統性發炎反應之動物模式……………………………………………………32
2. 大蒜精油之有機硫成分灌食……………………………………………………32
三、試藥成份及配製………………………………………………………………32
四、樣品收集………………………………………………………………………33
五、分析項目………………………………………………………………………34
六、統計分析………………………………………………………………………40
第三章、結果………………………………………………………………………41
一、灌食不同大蒜含硫成份對大鼠生長代謝的影響……………………………41
二、灌食不同大蒜含硫成份對系統性發炎大鼠臟器體重百分比的影響………41
三、灌食不同大蒜含硫成份對系統性發炎大鼠周邊血嗜中性球密度之影響…42
四、灌食不同大蒜含硫成份對於系統性發炎大鼠迴腸黏膜嗜中性球
浸潤指標MPO活性之影響 …………………………………………………42
五、灌食不同大蒜含硫成分對於系統性發炎大鼠血清中 sL-selectin
含量的影響……………………………………………………………………43

六、灌食不同大蒜含硫成分對於系統性發炎大鼠血清中 sICAM-1
含量的影響……………………………………………………………………44
七、灌食不同大蒜含硫成分對系統性發炎大鼠血清中 CINC-1含量的影響…44
八、灌食不同大蒜含硫成份對於嗜中性球表面黏附分子CD11b和CD18
表現之影響……………………………………………………………………45
九、灌食不同大蒜含硫成份對於小腸黏膜上皮層與完整小腸組織中黏附因子
ICAM-1、趨化因子CINC-1和促發炎激素TNF-α及IL-1β含量之影響…46
十、以MPO化學染色法觀察灌食不同大蒜含硫成份對於系統性發炎大鼠
迴腸組織MPO之影響…………………………………………………………47
十一、灌食不同大蒜含硫成份對於系統性發炎反應之大鼠迴腸組織完整性
之影響………………………………………………………………………47
第四章、討論………………………………………………………………………61
第五章、結論………………………………………………………………………67
第六章、參考文獻…………………………………………………………………68


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