阿托伐他汀作用於慶大黴素誘導大鼠腎毒性之差異性蛋白質體分析

Taipei Medical University Institutional Repository > 藥學院 > 臨床藥物基因體學暨蛋白質體學碩士學位學程 > 博碩士論文 > Item 987654321/59320

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題名:	阿托伐他汀作用於慶大黴素誘導大鼠腎毒性之差異性蛋白質體分析 Differential proteomic analysis of the effect of atorvastatin on gentamicin-induced nephrotoxicity in rats
作者:	李梅君 Lee, Mei-Chun
貢獻者:	李仁愛
關鍵詞:	阿托伐他汀 (atorvastatin);慶大黴素 (gentamicin);腎毒性;蛋白質體 atorvastatin;gentamicin;nephrotoxicity;proteomic
日期:	2015-06-17
上傳時間:	2020-08-11 15:46:09 (UTC+8)
摘要:	降血脂藥物阿托伐他汀 (atorvastatin, ATO)是3-hydroxy-3-methyl-3-glutarylcoenzyme A (HMG-CoA) reductase的抑制劑，它同時可以抑制non-steroidal isoprenoid compounds的合成，故具有多效性。研究發現ATO具有防止慶大黴素 (gentamicin, GM)引起的腎毒性，但詳細機轉仍不明，且GM引起腎細胞毒性的詳細機轉仍有待進一步闡明。因此，本研究藉由蛋白質體分析方法研究ATO對於GM引起腎毒性大鼠的療效影響與作用機轉。蛋白質體分析方法主要是將大鼠的腎臟組織均質化並利用螢光衍生化試劑DAABD-Cl後，經螢光高效能液相層析儀分離並收集表現量具有統計差異的波峰，再進入液相層析串聯質譜儀與MASCOT資料庫進行蛋白質的鑑定，再使用生物醫學分析研發軟體暨資料庫Ingenuity Pathways Analysis，建構蛋白質的毒性與功能與蛋白質之間的相關性路徑與機轉。蛋白質體分析結果，共有49個具有表現差異的蛋白質，GM腎毒性與粒線體功能失調、脂肪酸代謝與氧化壓力及其上游調控蛋白PPARα具高度相關性。與GM腎毒性最具相關的蛋白質有sodium-hydrogen exchanger regulatory factor (SLC9A3R1)、cathepsin V (CTSV)與transthyretin，蛋白質表現量在SLC9A3R1與transthyretin是下調，CTSV是上調。ATO介入治療下，蛋白表現量皆有呈現回復的趨勢，特別是CTSV與SLC9A3R1 ( p＜0.05)。由於CTSV與SLC9A3R1分別為溶酶體磷脂質代謝與細胞膜運輸的相關蛋白質，因此推測ATO可能是因為藉由改善磷脂質的代謝而降低phospholipidosis，以及增強調控細胞酸鹼值與容積的平衡來改善GM引起的腎毒性。另外，transthyretin與cathepsin V可能是GM引起腎毒性專一的biomarkers。 Atorvastatin (ATO) belonged to 3-hydroxy-3-methyl-3-glutaryl coenzyme A (HMG-CoA) reductase inhibitor. It also inhibited the synthesis of non-steroidal isoprenoid compounds and possessed pleotric effect. ATO role in preventing gentamicin (GM)-induced renal injury was well-established. However, the detailed mechanism remained obscure. Althought underlying multifaceted mechanisms involving in GM-induced nephrotoxicity were well known, further work on elucidating the major mechanism was needed. By using proteomic approach, we conducted mass spectrometry analysis to investigate the effects and mechanisms of ATO treatment in GM-induceed nephrotoxicity rats. The homogenate of kidney of rats were reacted with DAABD-Cl. The derivatized proteins were separated and quantified by high performance liquid chromatography with fluorescence detection (FD-HPLC). The significantly differential proteins were identified by liquid chromatography tandem mass spectrometry (LC-MS/MS) using a MASCOT database searching system. Finally, we used Ingenuity Pathway Analysis to find the relationships, mechanisms, functions, pathways of relevance, and protein networks. According to the experimental results, 49 proteins were significantly differential and identified. The most significant mechanisms of nephrotoxicity caused by GM were mitochondrial dysfunction, fatty acid metabolism and oxidative stress. Their upstream regulator was also found to be PPARα. The proteins involving in GM nephrotoxicity were sodium-hydrogen exchanger regulatory factor (SLC9A3R1), cathepsin V (CTSV) and transthyretin. The expression of CTSV was up-regulated and SLC9A3R1 and transthyretin were down-regulated in GM group. After ATO intervention, we observed a reverse enrichment pattern of the expression of them, especially in CTSV and SLC9A3R1 (p＜0.05). CTSV and SLC9A3R1were associated with lysosomal phospholipid metabolism and cell membrane transporters separately. Therefore, we predicted ATO may improve abnormal phospholipid metabolism and phospholipidosis caused by GM. ATO may also alleviate cell volume homeostasis and reverse the interference of GM to the transporter. Proteomic results also provided clues for GM-induced nephrotoxicity biomarkers such as CTSV and transthyretin.
描述:	碩士指導教授:李仁愛委員:田履黛委員:陳世銘
資料類型:	thesis
顯示於類別:	[臨床藥物基因體學暨蛋白質體學碩士學位學程] 博碩士論文

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