臺灣博碩士論文加值系統

活性氧物質會影響體內細胞的恆定，生產過多通常會與細胞死亡、疾病、以及老化有關。 老化會導致多個系統產生漸進性的功能損失，其中老年性聽力損傷也被認為與活性氧物質在內耳所引發的損傷有關。 此外，活性氧物質的大量形成也是噪音性聽力損傷的主要作用機制。耳蝸相較於其他器官更易受到氧化壓力的傷害，因為其內部構造的機械性感覺毛細胞在聲音刺激反應下有高度的能量代謝需求。 我們最近的研究發現，在具有聽損的漢丁頓氏症患者和小鼠模型中，腦型肌酸激酶的調控異常是造成聽損的重要原因，藉由肌酸飲食的補充則可提升耳蝸內腦型肌酸激酶的表現並減少氧化壓力，也藉此恢復小鼠的部分聽力。 因此，在我的研究中，我們假設腦型肌酸激酶的調控異常不單是造成漢丁頓氏症的聽損而已，可能也與其他形式的聽損有所關連。 我們希望驗證腦型肌酸激酶在活性氧物質所涉及的聽損傷害中扮演著重要的角色。實驗結果證實小鼠耳蝸中腦型肌酸激酶會隨著老化或噪音暴露後降低其表現量；給予肌酸的補充則可以提昇小鼠耳蝸腦型肌酸激酶的表現。 此外，在細胞培養的實驗中我們也證實了肌酸確實可以保護細胞減少活性氧物質的傷害以及其抗氧化的角色。

The generation of reactive oxygen species (ROS) will affect cellular homeostasis and high level of ROS are believed to be associated with cell death, disease, and aging. Aging results in progressive functional losses crossing multiple systems. Aging-induced hearing loss (AHL) may be contributed by ROS damage in the inner ear. In addition, the molecular mechanism for noise-induced hearing loss (NIHL) also involves the ROS formation. Cochleae are more vulnerable to oxidative stress compared to other organs because of the highly metabolic demands on their mechanosensory hair cells in response to sound stimulation. We recently showed that patients and mice with Huntington’s disease (HD) have hearing impairment and that the dysregulated phosphocreatine (PCr)‐creatine kinase (CK) system may account for this auditory dysfunction. Dietary creatine supplements have been shown to rescue the expression of cochlear brain type creatine kinase (CKB) in HD mice by reducing oxidative stress and thereby restoring their hearing. Thus, in my research, we hypothesize that dysregulated CKB not only be responsible for HD-related hearing impairment, but may also be implicated in several hearing impairments. We would like to investigate whether CKB plays an important role in ROS resultant hearing damages. Our results demonstrate that the expression of CKB in the cochlea would be reduced following the noise exposure or aging. Creatine supplementation helps in upregulating the expression of CKB in the mice cochleae. In addition, we showed that creatine, as an antioxidant, was demonstrated to protect auditory cells from ROS damage in vitro.

正文目錄 I
圖目錄 IV
中文摘要 V
ABSTRACT VI

第一章 緒論
第一節 活性氧化物質 1
第二節 腦型肌酸激酶 2
第三節 耳蝸之構造及功能 4
第四節 老年性聽損 5
第五節 噪音對耳蝸之影響 7
第六節 實驗動機與目的 8
第二章 材料與方法
第一節 試劑 10
第二節 動物模式 12
第三節 聽性腦幹反應 13
第四節 噪音模式 14
第五節 耳蝸組織樣本製備 14
第六節 石蠟組織切片 15
第七節 免疫組織化學染色 16
第八節 蛋白質定量 18
第九節 西方墨點法 19
第十節 細胞株培養 21
第十一節 模擬ROS環境 21
第十二節 胞內ROS偵測 22
第三章 結果
第一節 老化對腦型肌酸激酶表現的影響 23
第二節 噪音暴露後對腦型肌酸激酶表現的影響 25
第三節 肌酸補充對噪音暴露後肌酸激酶表現的影響 26
第四節 肌酸補充對細胞株之存活影響 28
第五節 肌酸補充對細胞內氧化壓力之影響 29
第四章 討論
第五章 結論
第六章 參考文獻

圖目錄
圖1 47
圖2 48
圖3 49
圖4 49
圖5 49
圖6 49
圖7 49
圖8 49
圖9 49
圖10 49

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