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系統識別號 U0026-0812200911425020
論文名稱(中文) 長期慢性運動對ApoE基因剔除小鼠胸主動脈血管之細胞凋亡相關蛋白表現的影響
論文名稱(英文) Effects of Chronic Exercise on the Expression of Apoptosis-Related Proteins in Aortas of ApoE-/- mice
校院名稱 成功大學
系所名稱(中) 生理學研究所
系所名稱(英) Department of Physiology
學年度 93
學期 2
出版年 94
研究生(中文) 邱怡珍
研究生(英文) Yi-Chen Chen
電子信箱 greenguava0824@yahoo.com.tw
學號 s3692105
學位類別 碩士
語文別 中文
論文頁數 47頁
口試委員 口試委員-江美治
指導教授-陳洵瑛
召集委員-任卓穎
中文關鍵字 細胞凋亡  動脈粥狀硬化  運動訓練 
英文關鍵字 apoptosis  chronic exercise  atherosclerosis 
學科別分類
中文摘要   動脈粥狀硬化是工業化社會中的主要致死原因之一。運動已廣泛的被認為具有治療動脈粥狀硬化的功用。在其他的研究指出,長期的運動訓練可以降低骨骼肌細胞凋亡的情形和降低促細胞凋亡蛋白的表現。前人的研究證明,在動脈粥狀硬化的損傷區上有細胞凋亡和促細胞凋亡相關蛋白表現的情形,而細胞的凋亡會導致動脈粥狀硬化的惡化。然而,有關長期的運動訓練,對於動脈粥狀硬化血管之細胞凋亡相關蛋白表現,仍是不清楚的。因此本篇論文的目的,是要探討長期的運動訓練,對於血管細胞上的細胞凋亡相關蛋白表現的影響。採用ApoE基因剔除小鼠餵食0.15% 高膽固醇飼料當作動脈粥狀硬化的動物模式,給予ApoE基因剔除公鼠跑步機的運動訓練,每天兩次,一次60分鐘 (速度為每分鐘15公尺),每週5天,持續十二週。結果發現,在給予運動訓練後,運動組的檸檬酸合成酵素活性明顯較控制組高,而體重明顯較控制組低。由oil red-O染色的結果顯示,運動訓練可降低胸主動脈竇上脂肪堆積的情形達18%,但不影響血清中總膽固醇和低密度脂蛋白膽固醇的含量。利用dihydroethidium的螢光染色可知,運動訓練後,運動組和控制組血管上超氧化物的表現沒有差異。以西方墨點雜合法測蛋白表現量,發現長期的運動訓練對血管細胞Bcl-2、超氧歧化酵素和熱休克蛋白72的表現沒有影響。綜合以上的結果可知,長期的運動訓練可以延緩動脈粥狀硬化的情形,且這可能和細胞凋亡相關蛋白的表現無直接的關係。
英文摘要  Atherosclerosis is the leading cause of mortality in modern society. It has been reported that chronic exercise is an anti-atherosclerotic therapy. Previous studies have demonstrated that chronic exercise training reduces the apoptosis and the expression of pro-apoptotic proteins in skeletal muscle cells. It is also noticed that apoptotic events are involved in atherosclerotic progression. Whether exercise training reduces apoptosis of vascular cells in atherosclerosis remains unknown. The objective of this study was to investigate the effects of chronic exercise training on the expression of the apoptosis-related protein in atherosclerotic aorta. Male ApoE-/- mice fed with 0.15% cholesterol diet were trained on a treadmill for 60 minutes twice per day (at speed of 15m/min), 5 days per week for 12 weeks. After exercise training, the citrate synthase activity was higher and the body weight was lower in the exercise group than in the control group. Results of oil red-O staining showed that exercise training reduced the fat deposits up to 18%. However, there were no significant differences in serum total cholesterol and low density lipoprotein cholesterol levels. By detecting the dihydroethidium fluorescence, the superoxide production between two groups was no difference. The expressions of the apoptotic regulating proteins were further determined by Western blotting. After exercise training, there were no differences in the expression of Bcl-2, Cu/Zn SOD and Hsp72 protein. In conclusion, chronic exercise training reduces the progression of atherosclerosis. This reduction may not relate to expression of apoptosis-related proteins.
論文目次 目錄
圖錄 v
表錄 vi
中文摘要 1
Abstract 2
緒論 4
(一) 動脈粥狀硬化與細胞凋亡的關係 4
(二) 細胞凋亡 4
(三) 氧化壓力 7
(四) 超氧歧化酵素 (superoxide dismutase,SOD) 8
(五) 熱休克蛋白質72 (heat shock protein 72,Hsp72) 9
(六) 運動 10
材料與方法 12
實驗動物 12
實驗動物的基因定型分析 (Genotyping) 12
高膽固醇飼料餵食 13
慢性中度運動 (Chronic Moderate Exercise) 13
動物犠牲 (Sacrifice)及組織取樣 13
血膽固醇之測定 15
冷凍切片 15
超氧化物的檢測 16
血管切片的脂肪堆積染色與量化 17
檸檬酸合成酵素活性 (Citrate Synthase Activity) 之檢測 17
蛋白質之萃取 19
蛋白質濃度之測定 19
蛋白質電泳與西方墨點雜合法 20
製備SDS-PAGE 20
電泳 (electrophoresis) 21
蛋白質電泳轉漬 (transfer) 21
雜交與偵測 (hybridization and detection) 21
統計分析 22
結果 23
基因定型分析結果 23
運動訓練後體重之變化 23
運動訓練後比目魚肌內檸檬酸合成酵素活性之變化 24
運動訓練後血清中血脂濃度之變化 24
運動訓練後血管上脂肪堆積之變化 25
運動訓練後血管上超氧化物表現之變化 25
運動訓練後血管上細胞凋亡相關蛋白表現之變化 25
討論 26
附錄 39
Krebs-Ringer-Hepes solution 39
6X Sample buffer 39
10X Running buffer 40
10X Transfer buffer 40
1X Transfer buffer 40
10X PBS 40
1X PBS-T 41
參考文獻 42
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