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研究生:胡瓊文
論文名稱:白血病病人周邊血及骨髓細胞中各種抗氧化酵素活性以及脂質過氧化物變化情形之探討
論文名稱(外文):The study of antioxidant enzymes and lipid peroxidation products in peripheral blood and bone marrow from leukemia patients
指導教授:張均昌蔡麗玉蔡麗玉引用關係
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
中文關鍵詞:白血病自由基
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我們將探討白血病病人其血液及骨髓細胞中SOD、Glutathione peroxidase、Glutathione reductase等酵素活性是否有變化,TAS的濃度以及脂質過氧化的程度。
檢體來源為收集高雄醫學大學附設中和紀念醫院血液腫瘤科之36例AML、24例ALL及31例CML共91例白血病病人周邊血之白血球、紅血球及血清,以及其骨髓細胞檢體;並同時以12位正常人做為控制組,收集其周邊血之白血球、紅血球及血清。
實驗結果發現:AML病人白血球之SOD活性與CuZn SOD活性與控制組相比,皆為有意義的下降 (p<0.05);而ALL及CML病人之SOD活性與CuZn SOD活性與控制組相比,並無有意義的差異。AML、ALL及CML病人之Mn SOD活性與控制組相比,都有增加的現象,其中ALL病人之Mn SOD活性為有意義的上升 (p<0.05)。AML、ALL及CML病人之脂質過氧化程度與控制組相比,都有下降的現象,但只有CML病人之脂質過氧化下降的程度有意義 (p<0.05)。
AML、ALL及CML病人都處於貧血狀態,因此其血紅素與控制組相比,都為有意義的降低 (p<0.001)。AML、ALL及CML病人紅血球之CuZn SOD、GPx、GRx等酵素活性與控制組相比,都是有意義的上升 (p<0.05)。
AML、ALL及CML病人血清之GRx酵素活性與控制組相比,都是有意義的上升 (p<0.05)。AML、ALL及CML病人之TAS濃度與控制組相比,都是有意義的下降 (p<0.05)。AML、ALL及CML病人之脂質過氧化程度與控制組相比,除CML為有意義的下降外 (p<0.05),其餘都沒有有意義的差異。
ALL及CML病人骨髓細胞中之SOD酵素活性與AML病人相比,都有明顯的上升,其中CML病人為有意義的上升 (p<0.05)。ALL及CML病人之CuZn SOD酵素活性與AML病人相比,都為有意義的上升 (p<0.05)。ALL及CML病人之TAS濃度與AML病人相比都是較低的,其中CML的降低在統計上是有意義的 (p<0.05)。
ALL病人白血球中之SOD與CuZn SOD活性,以及血清中GPx活性與AML病人相比都是較高的,且在統計上是有意義的 (p<0.05)。CML病人白血球中之SOD與CuZn SOD活性,與AML病人相比都是較高的,且在統計上是有意義的 (p<0.05)。
AML病人周邊血與骨髓細胞中SOD、CuZn SOD、Mn SOD、GPx酵素活性;TAS濃度以及脂質過氧化程度的比較,在統計上並無任何意義。ALL病人周邊血與骨髓細胞中SOD、CuZn SOD、Mn SOD、GPx酵素活性;TAS濃度以及脂質過氧化程度的比較,在統計上並無任何意義。CML病人周邊血與骨髓細胞中SOD、CuZn SOD、Mn SOD、GPx酵素活性;TAS濃度以及脂質過氧化程度的比較,在統計上並無任何意義。
由這些實驗的結果,我們得到的結論是:因為自由基、活性氧的產量增加、白血球、紅血球及血清的抗氧化酵素活性發生變化,而脂質過氧化的產物並無增加,顯示這些抗氧化酵素仍提供一定程度的保護,因此我們認為白血病病人的體內正面臨於mild oxidative stress的階段。但是白血球的噬菌能力與自由基的產生有關,因此自由基、活性氧的產量持續增加可能會對白血球的噬菌能力造成影響,對血液循環系統可接觸到的所有細胞如紅血球造成傷害,抑制這些細胞的正常功能,甚至造成這些細胞的老化、死亡。而GPx的活性會受Superoxide anion所抑制,SOD的活性則會被H2O2抑制,因此當活性氧的產量持續增加,可能會對這些抗氧化酵素造成銷耗殆盡或是完全抑制這些酵素的活性,造成細胞處與氧化的狀態,造成細胞功能不正常、結構的完整性失去,導致慢性發炎、癌化或是細胞大量死亡的發生。因此當自由基、活性氧的產量持續增加,可能會對這些抗氧化酵素造成銷耗殆盡或是完全抑制這些酵素的活性,造成細胞面對活性氧化物的攻擊而失去功能、癌化或是死亡。
I. 中文摘要…………………………………………………………………...1
II. 英文摘要………………………………………………………………….4
III. 緒論……………………………………………………………………...7
IV. 材料……………………………………………………………………...16
V. 方法……………………………………………………………………...18
A. 抗氧化酵素活性之測定方法…………………………………………...19
一. Superoxide dismutase (SOD) activity的測定………………………...19
二. Glutathione Peroxidase (GPx) activity的測定………………………...23
三. Glutathione Reductase (GRx) activity的測定………………………...26
四. Total Antioxidant Status (TAS) activity的測定………………………..28
五. Protein Assay (蛋白質分析) …………………………………………..30
B. 脂質過氧化產物之測定………………………………………………...33
一.血清TBA測定法……………………………………………………...33
二.白血球TBA測定法…………………………………………………34
C. 資料分析…………………………………………………………………36
VI. 結果………………………………………………………………………37
一. 各項實驗之control值……………………………………………………38
二.白血病病人與控制組白血球內各種抗氧化物表現與
脂質過氧化程度之比較…………………………………………………38
三. 白血病病人與控制組紅血球內各種抗氧化物表現之比較………………38
四. 白血病病人與控制組血清各種抗氧化物表現與脂質過氧
化程度之比較…………………………………………………………39
五. 白血病病人骨髓細胞中各種抗氧化物表現與脂質過氧化程
度之比較………………………………………………………………39
六. AML、ALL及CML病人白血球、紅血球及血清中各種抗
氧化物表現與脂質過氧化程度之比較……………………………………40
七. AML病人周邊血與骨髓細胞中各種抗氧化物表現與脂質過氧
化程度之比較……………………………………………………………40
八. ALL病人周邊血與骨髓細胞中各種抗氧化物表現與脂質過氧
化程度之比較……………………………………………………………40
九. CML病人周邊血與骨髓細胞中各種抗氧化物表現與脂質過氧
化程度之比較……………………………………………………………41
VII. 討論……………………………………………………………………42
VIII. 圖表……………………………………………………………………47
IX. 參考資料…………………………………………………………………78
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