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研究生:許德榮
研究生(外文):Dur-Zong Hsu
論文名稱:芝麻油與芝麻酚在治療敗血症大鼠之功效及其作用機轉
論文名稱(外文):Efficacy and Mechanism of Sesame Oil and Sesamol in Treating Sepsis in Rats
指導教授:劉明毅劉明毅引用關係
指導教授(外文):Ming-Yie Liu
學位類別:博士
校院名稱:國立成功大學
系所名稱:基礎醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:英文
論文頁數:175
中文關鍵詞:芝麻油芝麻酚內毒素盲腸結紥穿刺氧化性壓力脂質過氧化細胞激素內毒素結合蛋白死亡率
外文關鍵詞:endotoxinsesamolsesame oilcecal ligation and punctureoxidative stresscytokinelipid peroxidationmortalityLPS-binding protein
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  • 收藏至我的研究室書目清單書目收藏:1
  敗血症主要是由細菌的內毒素所引起,而且死亡率相當高,為台灣第十三大死亡原因。根據統計,在美國每年約有超過二十萬人死於敗血症,但是到目前為止,對於敗血症並沒有任何有效的治療方式。芝麻油為一種常用的保健食用油,在飼料中添加芝麻油長期餵食實驗動物,可降低由內毒素誘發之初發炎細胞激素,減少經盲腸結紥穿刺所引發敗血症動物之死亡率,然而其保護作用之機轉並不清楚。本研究之目的為探討芝麻油所含之木質素芝麻酚,對實驗性敗血症之保護作用及其可能機轉。本研究採用二個常用的動物模式,一個是直接給定量的內毒素來誘導敗血症,另一個則是對實驗動物作盲腸結紥穿刺手術,使動物產生腹膜炎而導致敗血症。實驗結果發現在內毒素誘導大白鼠的敗血症中,單一劑量芝麻油,無論是事前或事後給予,均能明顯降低脂質過氧化反應及體內氧化自由基的產生,並對內毒素所誘發的多器官衰竭具有保護作用,進而提高實驗動物的存活率;然而,給予礦物油及玉米油則無類似的保護作用。芝麻油中的抗氧化成份芝麻酚能降低盲腸結紥穿刺動物血清中之脂質過氧化、一氧化氮及介白質1beta的產生、改善多器官衰竭,且在腹膜炎發生事前或事後,給予芝麻酚能提高實驗動物的存活率。芝麻酚能減輕內毒素所誘發的氧化性壓力、降低血液氫氧自由基、peroxynitrite、超陰氧離子及一氧化氮的生成,減少腫瘤壞死因子alpha及介白質1beta的生成,降低內毒素誘發巨噬細胞中CD14及Toll-like receptor 4的表現,抑制內毒素與內毒素結合蛋白的結合。綜合以上結果,芝麻酚可能藉由抑制細菌內毒素與內毒素結合蛋白的結合,影響初發炎細胞激素的產生及自由基的生成,來減輕敗血症動物之多器官衰竭,降低敗血症所引起的死亡率。總之,本研究發現芝麻油及芝麻酚能有效治療實驗動物之敗血症,然未來在臨床應用來治療敗血症病人的效能,仍需進一步加以探討。
 Sepsis, mainly caused by bacterial infection, is one of the major causes of death in clinical patients. More than 200,000 patients die of sepsis every year in the United States, and is the top 13 cause of death in Taiwan. However, no effective interventions in treating sepsis have been reported during past two decades. Sesame oil is regarded as a health food. Sesame oil-supplemented diet decreases pro-inflammatory cytokines and mortality in septic mice. The aim of the present study was to examine the effects and the possible mechanisms of sesamol (3,4-methylenedioxyphenol), a lignan in sesame oil, against sepsis. Two common septic models were used, lipopolysaccharide (LPS) and cecal ligation and puncture (CLP). Data showed that a single dose of ses-ame oil can attenuate multiple organ failure and decrease mortality by reducing oxida-tive stress when given before and after the onset of sepsis in rats; however, both min-eral oil and corn oil did not show the similar effects. Sesamol decreased serum lipid peroxidation, nitric oxide, and IL-1beta production, and attenuated CLP-induced multi-ple organ injury. Furthernire, sesamol showed the proventive and therapeutic effects on mortality induced by CLP in rats. Sesamol significantly attenuated LPS-induced oxidative stress and multiple organ failure, reduced the generation of hydroxyl radical, peroxynitrite, and superoxide anion. Sesamol also inhibited nitric oxide production and inducible nitric oxide synthase expression, and the production of tumor necrosis factor-alpha and interleukin-1beta. Sesamol decreased the LPS-induced expression of CD14 and Toll-like receptor 4 in RAW264.7 cells. Sesamol potently decreased the binding between LPS and LPS-binding protein (LBP). We concluded that sesame oil attenu-ated multiple organ failure and decreased mortality in septic rats. Sesamol might be an important component of sesame oil for treating sepsis by inhibiting the binding be-tween LPS and LBP. However, more investigations are needed before application of sesamol in treating septic patients.
Abstract in Chinese..........................................6
Abstract.........................................7
Figure Content...................................9
Abbreviation....................................12
1. Introduction……………………………………………………15
1.1. Sepsis…………………………………………………………15
1.2. Oxidative stress in sepsis………………………………16
1.3. Pro-inflammatory cytokines………………………………20
1.4. Signal pathway of LPS-induced cytokine release……21
1.5. Sesame oil……………………………………………………22
1.6. Sesamol………………………………………………………22
1.7. The aims of this study……………………………………23
2. Experimental Design…………………………………………24
2.1. Effects of sesame oil on LPS-treated endotoxemia in rats…24
2.2. Protective effects of sesamol against sepsis in rats………27
2.3. Assessment of the effects of sesamol against endotoxemic rats...27
2.4. In vitro study of the effect of sesamol on the binding capacity of LPS to LBP…………………………………………………34
3. Materials and Methods…………………………………………35
3.1. Materials………………………………………………………35
3.1.1. Animals…………………………………………………35
3.1.2. RAW264.7 cells………………………………………35
3.1.3. Chemicals………………………………………………35
3.1.4. Antibodies………………………………………………38
3.1.5. ELISA assay kits………………………………………38
3.1.6. Expendables……………………………………………39
3.1.7. Equipments……………………………………………..39
3.1.8. Computer software…………………………………….41
3.2. Methods………………………………………………………...41
3.2.1. Blood collection…………………………………………41
3.2.2. Measurement of serum lipid peroxidation level……..41
3.2.3. Measurements of hydroxyl radical, superoxide anion, and peroxynitrite………………………………………42
3.2.4. Measurement of superoxide dismutase activity………43
3.2.5. Measurement of catalase activity……………………..44
3.2.6. Measurement of glutathione peroxidase activity…….45
3.2.7. Blood biochemistry……………………………………. 46
3.2.8. Histological evaluation of organ injury…………47
3.2.9. Measurement of serum nitrite concentration………48
3.2.10. Measurement of cytokines in serum…………………48
3.2.11. Leukocyte isolation……………………………………50
3.2.12. Western Blot………………………………………….. 51
3.2.13. LPS-LBP binding assay……………………………… 54
3.2.14. Cecal ligation and puncture (CLP)…………………56
3.2.15. Statistical analysis……………………………………56
4. Results……………………………………………………………58
5. Discussion………………………………………………………67
6. Conclusion………………………………………………………73
7. References………………………………………………………74
Table & Figures……………………………………………………86
Acknowledgement……………………………………………………109
Publication List…………………………………………………110
Publications………………………………………………………113
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