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研究生:孫婉寬
研究生(外文):Sun Wan Kuan
論文名稱:慢性缺氧能減緩內毒素所誘發的腎臟氧化性壓力
論文名稱(外文):Chronic hypoxia attenuates the endotoxin-induced renal oxidative stress
指導教授:陳朝峰陳朝峰引用關係鄭劍廷
指導教授(外文):Chen Chau FongChien Chiang Ting
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:生理學研究所
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:2003
畢業學年度:90
語文別:中文
論文頁數:78
中文關鍵詞:內毒血症活性氧物質缺氧預處理
外文關鍵詞:endotoxemiareactive oxygen specieshypoxia preconditioning
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內毒血症(Endotoxemia)可能會誘發活性氧物質(reactive oxygen species),進而促使組織器官產生功能障礙等問題。在許多臨床的報告也發現內毒素易造成患者急性腎衰竭。過去我們實驗室發現事先給予腎臟一段時間的缺氧預處理後(hypoxic preconditioning),腎臟在遭受到缺血再灌流傷害的耐受力有增加的情形。不過,低氧預處理對於內毒素誘發的氧化性傷害,其在腎臟的保護效果詳細機制仍未明。我們將比較給予內毒素之後,腎臟的氧化性傷害在海平面鼠(sea level groups, SL)及長期缺氧鼠(high altitude groups, HA; 將動物置放在模擬高度5,500 m的低壓低氧太空艙中,每天15小時,共放置4週)間的不同。首先將SL鼠及HA鼠分成未給予LPS (lipopolysaccharide)處理的控制組(SLCL, HACL)及由腹腔給予LPS 4 mg/kg 處理48小時後的LPS組(SLLPS, HALPS)。實驗過程中利用多頻道生理記錄儀記錄血流動力學的變化,並以化學冷光測定儀來偵測血液、活體腎臟表面及離體腎絲球中自由基釋出量的改變,最後利用西方墨點法測量腎臟皮質與髓質中抗氧化酵素Cu/Zn SOD蛋白質含量的變化。實驗結果顯示,給予LPS處理後SLLPS組及HALPS組的平均動脈壓低於控制組;但腎血流方面並無明顯的變化。在自由基釋出量方面,給予LPS處理後不論是血液、活體腎臟表面或是離體腎絲球中自由基的釋出量皆明顯的增加(SLLPS, HALPS);而HALPS 組的自由基釋出量明顯的少於SLLPS組。在Cu/Zn SOD的蛋白質表現變化方面,給予LPS處理的動物(SLLPS, HALPS) ,其Cu/Zn SOD蛋白質表現比控制組(SLCL,HACL)少。因此,我們推論低氧預處理對於內毒素所誘發的氧化性傷害有某種程度的保護效果,且抗氧化酵素可能是此保護機制中的重要因子之一。

Endotoxemia may induce the generation of reactive oxygen species (ROS), that may cause the dysfunction of tissues and organs. Our previous studies showed that the kidney after hypoxic preconditioning was more tolerant to the ischemia-induced oxidative injury. It is unclear whether the renal protective ability induced by hypoxic preconditioning can reduce endotoxin-induced injury. We compared the renal oxidative stress after endotoxin treatment (4 mg/kg, i.p. 48 h earlier) between control (SL) rats and chronic hypoxic (HA) rats, that were placed in a hypobaric chamber (380 Torr, 5,500m) 15 h/day for 4 weeks prior to experiment. The SL rats were randomized into two groups, one group is control group (SLCL), in the other group the rats received LPS(Lipopolysaccharide, from E.coli, Sigma, USA) 4mg/kg, ip, pretreat 48h (SLLPS). The HA rats randomized into two groups, one group is control group (HACL), in the other group the rats received LPS(Lipopolysaccharide, from E.coli, Sigma, USA) 4mg/kg, ip, pretreat 48h (HALPS). Homodynamic changes were recorded by multiple-channel record, and determined the amount of reactive oxygen species (ROS) from renal surface in vivo and from whole blood and isolated glomeruli in vitro by an ultrasenstive chemiluminscence method. We used the western blotting method to determine the protein expression of the Cu/Zn SOD and the Mn SOD from the renal cortex and renal medulla. Results showed that endotoxin significantly increased ROS release from kidney surface, whole blood and isolated glomeruli in the SLLPS groups and the HALPS groups. However, the enhanced oxidative stress was significantly reduced by the hypoxic pretreatment. The expression of Cu/Zn superoxide dismutase (Cu/Zn SOD) in SLLPS groups were less than HALPS groups. Our results suggest that HA evoked something might contribute to renal protection against endotoxin-induced oxidative stress, antioxidant may be play a role of the protect mechanism.

誌謝……………………………………………………………………...i
圖次…………………………………………………………………….iii
中文摘要………………………………………………………………..v
英文摘要………………………………………………………………vii
緒論及文獻回顧
一、 內毒素……………………………………………………….1
二、 缺氧………………………………………………………….6
三、 氧化性傷害…………………………………………………12
材料與方法
一、 實驗動物…………………………………………………….20
二、 各類插管…………………………………………………….20
三、 儀器設備…………………………………………………….21
四、 實驗藥品及來源…………………………………………….22
五、 藥品的配製………………………………………………….24
六、 一般手術過程……………………………………………….24
七、 實驗過程…………………………………………………….25
八、 動物活體腎臟內O2.-的偵測………………………………..30
九、 血液及離體腎絲球中O2.-的偵測………………………32
十、 西方墨點法………………………………………………….33
十一、 資料處理及統計方法……………………………………..35
實驗結果
一、 內毒素對腎功能的影響……………………………………36
二、 內毒素對腎臟內自由基釋出量的影響…………………….38
三、 內毒素處理後在SL組及HA組的腎皮質與髓質中抗氧化酵素Cu/Zn SOD與Mn SOD蛋白質表現量的比較………….40
討論……………………………………………………………………..42
結論……………………………………………………………………..49
參考文獻………………………………………………………………..64
附錄一、自由基與血液淨化學術研討會摘要…………………………77
附錄二、生物醫學聯合學術年會摘要………………………………..78
圖 次
圖一、活體化學冷光分析儀測量自由基之示意圖……………………53
圖二、化學冷光分析儀裝置圖…………………………………………54
圖三、體液膨脹對SLCL、HACL、SLLPS、HALPS組尿流量改變的比較……………………………………………………………..55
圖四、 體液膨脹對SLCL、HACL、SLLPS、HALPS組累積尿流量(aUO)改變的比較……………………………………………………..56
圖五、體液膨脹對SLCL、HACL、SLLPS、HALPS組絕對鈉離子排泄率的比較……………………………………………………..57
圖六、體液膨脹對SLCL、HACL、SLLPS、HALPS組累積鈉離子排除量的比較…………………………………………………….58
圖七、化學冷光分析儀測量血液中自由基釋出量之原始圖……….59
圖八、化學冷光分析儀測量離體腎絲球中自由基釋出量之原始圖…60
圖九、化學冷光分析儀測量活體腎臟表面自由基釋出量之原始圖…61
圖十、LPS處理後SL組及HA 組血液、離體腎絲球與活體腎臟表面自由基增加量之百分比……………………………………….62
圖十一、LPS處理後SLCL、HACL、SLLPS、HALPS組腎臟皮質及髓質抗氧化酵素Cu/Zn SOD 的蛋白質表現差異…………..63
附圖、LPS結構圖……………………………………………………..52

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