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系統識別號 U0026-0812200915202440
論文名稱(中文) 運動訓練對第一型糖尿病鼠於內毒素血症後腦部細胞激素和相關蛋白的影響
論文名稱(英文) Effects of exercise preconditioning on cytokines and related proteins in the brain of streptozocin-induced diabetic rat after systemic endotoxemia
校院名稱 成功大學
系所名稱(中) 物理治療研究所
系所名稱(英) Department of Physical Therapy
學年度 97
學期 2
出版年 98
研究生(中文) 張欣如
研究生(英文) Sin-ru Jhang
學號 t6696109
學位類別 碩士
語文別 英文
論文頁數 46頁
口試委員 指導教授-洪菁霞
口試委員-鄭瑞棠
口試委員-陳郁文
中文關鍵字 細胞凋亡    內毒素血症  運動  糖尿病 
英文關鍵字 endotoxemia  exercise  diabetes  brain  apoptosis 
學科別分類
中文摘要 背景及研究目的:隨著社會進步的變遷,糖尿病是成為影響世界最鉅的代謝性疾病。除了高血糖的症狀以外,慢性系統性低度發炎也是其特徵之一。第一型糖尿病主要由於免疫調節不全所造成,牽動巨噬細胞分泌免疫前驅物質,例如間白素-1β(Interleukin-1β, IL-1β)以及腫瘤壞死因子α(TNF-α)進而破壞胰島細胞。然而IL-1β需要經由caspase-1分解才有生物性活化。TNF-α則是會誘發細胞凋亡執行者caspase-3的活化。臨床上,糖尿病患容易受格蘭氏陰性菌細胞壁上的脂多醣體(lipopolysaccharide, LPS)感染,變成內毒素血症,動物實驗也發現發炎狀態與器官組織破壞有相關性。而腦部病變是最近十年來被認定為糖尿病的併發症之一,其細胞凋亡和免疫反應的相關性仍未定論。運動被認為有多效果的治療,也會增加腦部抗凋亡蛋白熱休克蛋白72(HSP72)。因此,本篇實驗在探討第一型糖尿病鼠腦部基礎細胞激素的量、罹患內毒素血症後其免疫系統的反應,以及運動對於腦部免疫功能的效果和凋亡與非凋亡蛋白的表現量。方法:選用八週大成熟雄性Wistar品系鼠,將其分為正常和糖尿病組。再細分為運動和靜態組。運動訓練的劑量為每週五次,每次30分鐘至一小時,速度為每分鐘20公尺到25公尺,為期一個月。並於後一次運動訓練24小時後注射LPS或是生理食鹽水。三小時後取下腦部組織分析。利用酵素結合免疫吸附法(Enzyme-linked immunosobent assay, ELISA)確定免疫前驅物質濃度,和西方墨點法(Western blotting)確認蛋白質的表現量。結果:在海馬迴和前額皮質IL-1β和TNF-α會受到LPS的影響。疾病狀態會影響NTS在TNF-α的表現量,其和LPS在下視丘有交互作用。運動和LPS對於IL-1β在下視丘有交互作用。此外,運動會影響正常大鼠的caspase-1在前額皮質的表現量,也會增加caspase-3在下視丘的表現。除了紋狀體和NTS以外,運動訓練皆會增加腦部HSP72的產生。結論:運動訓練雖然未能完全改變免疫前驅物質的表現量,也會增加正常大鼠凋亡蛋白的表現。然其增加抗凋亡蛋白HSP72的產生,有可能在腦部細胞凋亡佔有重要的角色,仍需更深入的瞭解。
英文摘要 Objective: Because of dysregulation of immune system, diabetic patients had higher rate to suffer from the infection and endotoxemia. Pro-inflammatory cytokines such as interleukin-1beta (IL-1β) was increased during endotoxemia. Interleukin-1beta converting enzyme (Caspase-1 or ICE) and tumor necrosis factor alpha (TNF-α) which induce apoptotic proteins (Caspase-3) were all leading to cells damage. In addition, diabetic rodents had showed higher hippocampal astrocytosis and cell loss. However, exercise may attenuate tissue damage by changng pro-inflammatory cytokines in plasma and inducing anti- apoptotic proteins, heat shock protein 72 (HSP72). The purpose of this study is to determine the effects of exercise on endotoxin-induced immune response, apoptotic and anti-apoptotic proteins expression in the brain of diabetic and normal rats. Materials and Methods: Adult male Wistar rats were treated with streptozotocin to destroy pancreatic β cells irreversibly. Normal and diabetic rats were randomly assigned to exercise and sedentary groups respectively. The exercise groups were trained to run on the treadmill 5 days/week, 30-60 min/day with intensity 20-25m/min for 4 weeks. Twenty-four hours after the last exercise session, rats received an intravenous injection of lipopolysaccharide (LPS) to induce systemic endotoxemia. At the 3rd hour after LPS injection, we determined the level of IL-1β, TNF-α, caspase-1, caspase-3 and HSP-72 in the rat brains including hippocampus, hypothalamus, frontal cortex, striatum, and nucleus tractus solitarius (NTS). Results: After administration of LPS, the TNF-α level was significant increased in hippocampus in normal and diabetic rats. The expression of pro-caspase-1 was increased by healthy condition but not influenced by LPS in hippocampus. The IL-1β level was significant increased in frontal cortex in normal and diabetic rats after LPS injection. There was an interaction between LPS treatment and healthy condition in IL-1β level in hypothalamus. Healthy condition had altered the expression of pro-caspase-3 in hypothalamus. The effect of healthy condition on changing TNF-α level was observed in NTS. The expression of pro-caspase-1 was significant higher increased with exercise pre-conditioning in normal rats’ frontal cortex. Compared exercise groups with sedentary ones, exercise did have effects on HSP72 expression in all brain area except striatum. Considered IL-1β level in hypothalamus, LPS treatment resulted in significant increasing in normal rats, and exercise would interact with LPS treatment in diabetic group. Conclusion: Diabetic rats were prone to apoptosis and inflammatory than normal ones. Exercise can induce protective proteins which alleviate brain cell damage in diabetic rats during systemic endotoxemia.
論文目次 Abbreviations-------------------------------------------------------------------------------------------I
Abstract-------------------------------------------------------------------------------------------------II
Chinese Abstract----------------------------------------------------------------------------------------------IV
Acknowledge----------------------------------------------------------------------------------------------------VI
List of figures---------------------------------------------------------------------------------------VIII
Introduction---------------------------------------------------------------------------------------------1
Methods and materials---------------------------------------------------------------------------------6
Results--------------------------------------------------------------------------------------------------11
Discussion---------------------------------------------------------------------------------------------14
References---------------------------------------------------------------------------------------------19
自述----------------------------------------------------------------------------------------------------46
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