在原核及真核細胞中，熱休克蛋白一直存在，但當溫度升高時，增加熱休克蛋白的合成是非常常見的現象。當細胞面臨壓力時會造成增加熱休克蛋白的合成，因此熱休克蛋白的誘導被認為是細胞壓力普遍的標記，與一個細胞損害的廣泛來源(包括溫度、UV、重金屬及能源耗盡等)有關。星蟲動物是兩側對稱、不分體節的海洋底棲動物，全球已報導的大約有300種，如土釘星蟲和方格星蟲。本研究，是以蛋白質體學的觀點，探討不同汙染程度下，星蟲抵抗環境汙染的機制。並以新竹香山及彰化王功濕地之方格星蟲為樣本，於野外環境中採樣，比較不同汙染程度下，方格星蟲之HSP70及HSP90表現之差異。結果顯示，新竹香山的多環芳香烴(PAHs)和有機氯農藥(OCPs)濃度無論在環境中或是方格星蟲內，均高於彰化王功的濃度。不論在新竹香山或是彰化王功\\\\其多環芳香烴(PAHs)及有機氯農藥(OCPs)濃度，在環境底泥中的含量均高於腸道中底泥含量及在身體組織中的含量。主成分分析結果指出，彰化王功及新竹香山方格星蟲體內的PAHs來源組成相似；而環境及腸道中底泥之PAHs其組成來源組成相近。比較兩地之HSP70及HSP90發現，HSP70在兩地的表現量並沒有顯著差異，而香山濕地的HSP90表現量則明顯高於王功濕地，顯示HSP90在對抗環境中的汙染可能佔有重要的角色，並可藉由偵測方格星蟲的HSP90表現量，作為監測環境變化之指標。

Synthesis of heat shock proteins (Hsps) in response to elevated temperatures and other denaturing agents (including UV, heavy metals, and energy depletion) is a common productive response of prokaryotic and eukaryotic cells. Therefore, increasing in production of heat shock protein has been considered a sign of cells under stress. The phylum Sipuncula comprises of about 300 species worldwide; they are bilaterally symmetrical, unsegmented, and deposit feeding marine worms common in the substrate of wetland. In this research, I studied the dominant species Sipunculus nudus and analyzed it proteomics to reveal how it responses to persistent organic pollution. In order to compare the difference between HSP70 and HSP90, Sipunculans were sampling from two wetland (Xiangshan and Wanggong) in Hsinchu and Changhua,, respectively. The results showed that the concentrations of PAHs and OCPs both in substrate and S. nudus tissues from Xiangshan were higher than Wanggong. The concentrations of PAHs and OCPs in both areas showed that the environmental- sediment levels were higher than the sediment inside the gut or the body tissues. Principal component analysis showed that the compositions in S. nudus from both area were similar; however PAHs compositions from the environmental sediment or the sediment of the gut were similar. When the concentrations of HSP70 and HSP90 between both areas were compared, HSP70 expressions did not significant differed in both areas, while HSP90 express was higher in the S. nudus from Xiangshan than Wanggong. For S. nudus Hsp90 was upregulated in highly polluted area (i.e. Xiangshan). I propose that tissue expression of HSP90 plays an important role in the survival of S. nudus, and detection of HSP90 may provide pollution information of the surrounding environment.

總目錄
謝辭...........................................................i
中文摘要.......................................................ii
英文摘要......................................................iii~ iv
總目錄.........................................................v~vii
表目錄.........................................................viii
圖目錄.........................................................viii
壹、前言.......................................................1~2
貳、文獻回顧...................................................3~13
2-1持久性有機汙染物...........................................3
2-1-1 多環芳香烴.............................................3
2-1-2 多環芳香烴的來源.......................................4
2-1-3多環芳香烴的危害........................................5
2-1-4 有機氯農藥.............................................5
2-1-5有機氯農藥之分類........................................6
2-1-6有機氯農藥之危害........................................6~7
2-2熱休克蛋白 .................................................8
2-2-1 HSP70..................................................8~9
2-2-2 HSP90..................................................9
2-3星蟲動物門..................................................10~13
2-3-1形態特徵................................................10
2-3-2生態特點................................................10
2-3-2-1生活型態............................................10~11
2-3-2-2 溫度範圍............................................11
2-3-3食性....................................................11
2-3-4生理....................................................11~12
2-3-5生殖和發育..............................................12
2-3-6裸體方格星蟲............................................12~13
2-3-6-1形態特徵............................................12~13
2-3-6-2生活環境............................................13
2-3-6-3地理分布............................................13
叁、研究方法....................................................14~24
3-1 採樣地點...................................................14
3-1-1 王功濕地的樣本來源.....................................14
3-1-2 香山濕地的樣本來源.....................................14
3-2 樣品前處理.................................................14
3-3 多環芳香烴及有機氯農藥之試藥與儀器設備.....................15~16
3-3-1 標準品.................................................15
3-3-2 試藥及器材前處理.......................................16
3-3-2-1 試藥前處理.........................................16
3-3-2-2 器材前處理.........................................16
3-4 實驗方法...................................................17~24
3-4-1 多環芳香烴及有機氯農藥分析流程.........................17~18
3-4-1-1 儀器分析...........................................18
3-4-1-2 定量方法...........................................19
3-4-1-3 多環芳香烴與有機氯農藥之品質保證與品質控制.........20
3-4-1-4 空白試驗...........................................20
3-4-1-5 方法偵測極限.......................................20
3-4-1-6 主成分分析.........................................21
3-4-2蛋白質凝膠電泳..........................................22
3-4-2-1蛋白質萃取.........................................22
3-4-2-2 蛋白質濃度定量分析.................................22
3-4-2-3一維聚丙烯胺凝泳..................................22~23
3-4-3 西方轉印法............................................23~24
3-4-4 統計方法..............................................24
肆、結果.......................................................25~28
4-1香山及王功濕地方格星蟲體長與體重之分析......................25
4-2香山及王功濕地多環芳香烴含量分析............................25
4-2-1脂肪含量分布............................................25
4-2-2香山及王功濕地底泥及方格星蟲蟲體之多環芳香烴含量分析....25
4-2-3香山及王功濕地方格星蟲脂肪中多環芳香烴之含量分析........26
4-2-4多環芳香烴之分布情況....................................26
4-3香山及王功濕地有機氯農藥含量分析............................27
4-3-1香山及王功濕地底泥方格星蟲蟲體之有機氯農藥含量分析......27
4-3-2香山及王功濕地方格星蟲脂肪中有機氯農藥之含量分析........27
4-4 香山及王功濕地方格星蟲之HSP70表現分析.....................28
4-5香山及王功濕地方格星蟲之HSP90表現分析......................28
伍、討論........................................................29~31
5-1 脂肪含量分布................................................29
5-2多環芳香烴之分布情況.........................................29
5-3香山及王功濕地方格星蟲之HSP70表現分析.......................30
5-4香山及王功濕地方格星蟲之HSP90表現分析......................31
陸、結論........................................................32
柒、參考文獻....................................................33~39
附錄............................................................55~57

表目錄
表一、21種常見之PAHs其分子量及結構式..........................40~41
表二、有機氯農藥的特性..........................................42
表三、香山及王功濕地分析樣品之多環芳香烴擬似標準品回收率........43
表四、分析樣品之多環芳香烴標準品回收率..........................43
表五、分析樣品之有機氯農藥標準品回收率..........................44
表六、多環芳香烴之偵測極限......................................45
表七、有機氯農藥之偵測極限......................................46

圖目錄
圖一、實驗採樣地.................................................47
圖二、多環芳香烴及有機氯農藥分析流程.............................48
圖三、新竹香山及彰化王功濕地的方格星蟲之平均脂肪含量(p < 0.05 :＊).49
圖四、新竹香山及彰化王功濕地之平均多環芳香烴含量比較(p < 0.05 :＊).50
圖五、新竹香山及彰化王功濕地方格星蟲脂肪中平均多環芳香烴之總濃度
(p < 0.05 :＊)...............................................50
圖六、香山及王功濕地多環芳香烴之主成分分析.......................51
圖七、新竹香山及彰化王功濕地15種多環芳香烴之不同環數組成分析....52
圖八、新竹香山及彰化王功濕地之平均有機氯農藥含量比較(p < 0.05 :＊).53
圖九、新竹香山及彰化王功濕地方格星蟲脂肪中平均有機氯農藥之總濃度
(p < 0.05 : ＊)..............................................53
圖十、新竹香山及彰化王功濕地方格星蟲HSP70及HSP90之表現量 ......54

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