臺灣博碩士論文加值系統

噪音是常見造成聽損傷害的原因之一，也是全世界最為常見的職業傷害，其病理成因包含活性氧化物質在耳蝸中的大量生成導致噪音聽損傷害的發生。此外，耳蝸內作為接收聲音的機械性毛細胞受聲音刺激後對能量具有高度新陳代謝的需求，使其對氧化壓力的傷害較其它組織器官而言更為敏感。先前的研究發現卡洛里限制下可以減少氧化傷害進而降低年老性聽損傷害機率; 進一步的的研究顯示去乙醯化酶sirt3對卡洛里限制下具有調節、降低氧化傷害的角色之一。有研究指出，在神經退化相關疾病中，腦型肌酸激酶表現與活性氧化物質及聽損傷害間具有關聯性。腦型肌酸激酶在神經組織中對能量調控占有重要的角色，我們實驗室過去研究指出在神經退化性的相關疾病中，增加腦型肌酸激酶的表現量可降低聽力損傷。因此本研究中我們假設卡洛里限制下可以減少噪音所生成的活性氧化物質，進而降低聽損傷害的發生，希望能驗證卡洛里限制對預防噪音性聽損的影響。實驗結果顯示卡洛里限制下實驗組的體重較正常飲食的對照組輕，且其去乙醯化酶sirt3與腦型肌酸激酶的表現量也較對照組高。40%熱量飲食控制組初步看起來對聽損傷害有保護效果。當細胞腦型肌酸激酶大量表現時會提升粒腺體的活性。且腦型肌酸激酶大量表現的細胞對維生素K3 menadione (2-methyl-1,4-naphthoquinone) 傷害教具有保護效果。

Noise is one of the most common causes of hearing loss, and one of the most common occupational illnesses in the world. Noise-induced hearing loss (NIHL) following cochlear damage to noise trauma was linked to a common pathogenesis involving the formation of reactive oxygen species (ROS). Cochleae are more vulnerable to oxidative stress than other organs because of the high metabolic demands of their mechanosensory hair cells in response to sound stimulation. Previous studies have shown that calorie restriction are able to reduce oxidative damage to prevent age-related hearing loss. Recent data suggests that Sirt3 plays a main role to mediate reduction of oxidative damage under calorie restriction. Previous studies shown that neurodegenerative genetic disorder related brain-type creatine kinase (CKB) is sensitive to oxidative modification. Overexpression of CKB might be able to restore their hearing. We therefore hypothesize that caloric restriction can also rescue the noise-exposed hearing damage. In this study, our specific aim is to investigate the impact of calorie restriction on hearing preservation in noise-exposed animals. Our preliminary results demonstrated that calorie restriction in mice resulted in weight loss and elevation of Sirt3 and CKB proteins. After noise exposure, animals treated with 40% calorie restriction diet for 12 weeks exhibited a better ABR thresholds as compared to the control. We also found that overexpression of CKB in HEI-OC1 cells would result in a better functional state of mitochondria and were more resistant to the cytotoxicity of menadione than the control.

中文摘要 V
Abstract VII
第一章 緒論 1
第一節 活性氧化物質 1
第二節 卡洛里飲食限制 2
第三節 去乙醯化酶sirt3 3
第四節 腦型肌酸激酶 4
第五節 耳蝸的功能與構造 4
第六節 噪音性聽損傷害 5
第七節 實驗動機與目的 5
第二章 材料與方法 7
第一節 試劑 7
第二節 動物模式 10
第三節 聽性腦幹反應 10
第四節 噪音模式 11
第五節 耳蝸組織樣本製備 11
第六節 石蠟組織切片 12
第七節 免疫組織化學染色 12
第八節 蛋白質定量 13
第九節 西方墨點法 14
第十節 細胞株培養 16
第十一節 模擬ROS環境 16
第十二節 胞內ROS偵測 16
第十三節 細胞轉染 17
第三章 結果 18
第一節 卡洛里限制對去乙醯化酶sirt3的影響 18
第二節 卡洛里限制對腦型肌酸激酶的影響 19
第三節 卡洛里限制下對噪音曝露的影響 20
第四節 氧化壓力環境對細胞株之影響 22
第五節 腦型肌酸激酶大量表現對細胞株的影響 23
第六節 腦型肌酸激酶大量表現細胞株對氧化傷害的保護效果 24
第四章 討論 26
第五章 結論 29
第六章 參考文獻 30

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