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標題: | 探討克林黴素對細胞自噬與APP水解的影響 Investigation of the Effect of Clindamycin on Autophagy and APP Processing |
作者: | Chwen-Yu Chen 陳淳于 |
指導教授: | 孔繁璐(Fan-Lu Kung) |
關鍵字: | 阿茲海默氏症,克林黴素,細胞自噬,APP 水解途徑,LC3-II, Alzheimer’s disease,Clindamycin,Autophagy,APP Processing,LC3-II, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 阿茲海默氏症(Alzheimer’s disease,AD)是一類廣受重視的漸進式神經退化性疾病,其在組織病理學的特徵之一為老年斑塊。在以往的研究指出,老年斑塊主要由具有細胞毒性的Aβ的堆積所形成的。而Aβ則是APP進行β processing,經由β-secretase和γ-secretase水解後所得的產物。Aβ的生成與堆積被認為與許多因素有關,其中之一是細胞自噬。
細胞自噬是細胞在處於挨餓環境下,藉由分解一些次要的蛋白質或胞器,以應付需要和維持細胞存活的機制。近年來科學家發現,細胞自噬不僅發生在細胞營養缺乏的情況,也會在受到外來刺激或輻射導致有胞器受損(如粒線體)而威脅細胞存活時觸發,能避免細胞進行細胞凋亡而有自我保護的功用。許多與阿茲海默氏症的相關的研究中指出,當誘發細胞自噬時,會造成APP 水解產物Aβ生成量減少以及幫助清除Aβ的堆積。至於細胞自噬如何影響Aβ的生成與堆積,甚至如何調控APP的水解途徑,目前仍然不是很明確。 本研究觀察在使用克林黴素(clindamycin)之下,對於GBM8401與SH-SY5Y兩種細胞株細胞自噬的影響。從西方墨點法的結果發現,GBM8401與SH-SY5Y中的LC3-II的量皆有明顯的上升,表示在克林黴素的作用下,兩種細胞株都有細胞自噬的發生。另外,本研究也觀察了在克林黴素作用下,APP 水解途徑是否受到改變。西方墨點法的結果顯示,在GBM8401中雖然無法看到C99的訊號,但是C83的結果可能有隨著克林黴素的作用而減少的趨勢。然而,也在GBM8401的細胞培養液中發現sAPPα可能有增加的現象。綜合上述結果,本研究初步觀察到克林黴素對於細胞自噬與APP水解途徑的影響。關於在克林黴素的處理之下,APP水解途徑會如何改變以及細胞自噬與APP水解途徑改變之間的關係,仍需要進一步的研究證明。 Alzheimer’s disease is a kind of progressive neurodegenerative diseases to which people paid much attention. One of the features of Alzheimer’s disease in histopathology is the formation of senile plaques. Early research has shown that senile plaques are formed from the aggregation of cytotoxic Aβ. Aβ is the product of amyloid precursor protein (APP) by β processing, which is cleaved by β-secretase and γ-secretase successively. Many studies have claimed that dozens of factors influence on the synthesis and clearance of Aβ, and one of which is autophagy. Autophagy is a way by which cells are able to degrade some proteins and organelles unnecessary to meet the need and survive under starvation. Recently, scientists found that autophagy is not only switched on when cells are starving, but also when cells are under stimulation or radiation that some organelles are damaged, such as mitochondria, and survival is threatened. By autophagy, cells can prevent themselves from apoptosis. Thus, autophagy is protective. In Alzheimer’s disease, some studies show that once autophagy is induced, it leads to decreasing synthesis of Aβ and clearance of Aβ aggregation. However, how autophagy work on the synthesis and clearance of Aβ, and even how autophagy work on the alternation of APP processing pathways remains unclear. In this study, two cell lines, GBM8401 and SH-SY5Y, are used to see if autophagy is regulated under clindamycin treatment. Western blot results show that the level of LC3-II increase obviously in GBM8401 and SH-SY5Y, indicating that clindamycin induces autophagy in two cell lines. Also, GBM8401 is used to see if APP processing pathway is changed under clindamycin treatment. Western blot results show that on one hand, in cell lysate C83 may tend to decrease. On the other hand, however, sAPPα seems to increase in conditioned medium. Above all, this study shows the preliminary effect of clindamycin on autophagy and APP processing pathways. Still, further studies are needed to investigate how clindamycin change APP processing pathway and the relationship between autophagy and APP processing pathways alternation. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7810 |
DOI: | 10.6342/NTU201700554 |
全文授權: | 同意授權(全球公開) |
顯示於系所單位: | 藥學系 |
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