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研究生(外文):Ya-Fan Wu
論文名稱(外文):Pathological Responses of Body Wall Collapse Induced by Cadmium in Earthworm Eisenia Andrei
外文關鍵詞:cadmiumantioxidantearthwormoxidative stressROSmuscleapoptosisnecrosis
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重金屬鎘(Cd)由於在工業上普及的應用,加上沒有良好管制而隨著工廠排放釋出於環境中。鎘透過食物鏈在生物體內大量累積,因而造成生物體全面性的傷害。蚯蚓分佈廣泛且族群量又大,再加上能適當地偵測土壤中的有毒物質,中毒後便能透過異常的生理狀況即時地反映出來,所以蚯蚓對於土壤汙染能成為一種良好的生物偵測器。在實驗中用愛氏蚓(Eisenia andrei)來研究高劑量鎘毒性所造成的毒理反應。蚯蚓在鎘處理後,表現出全身性的病徵,包括表皮皺褶、充血、滲血、循環不佳等。除此之外,蚯蚓表現出一個特殊的病理現象─體壁不正常凹陷。在實驗中,利用切片技術,觀察到蚯蚓在鎘處理後所產生的體壁凹陷,是由肌肉組織大規模崩解所造成。另外,鎘處理之後,使蚯蚓組織中之H2O2濃度顯著增加,並且抗氧化酵素catalase的活性亦顯著降低。若在處裡鎘的蚯蚓同時處理各種抗氧化劑(EUK134、薑黃素、迷迭香酸、兒茶素、維他命C ),組織內的鎘累積量會大幅降低,而死亡率也會顯著降低,除此之外,所觀察到之病徵比例也大幅降低,表示抗氧化劑確實能保護蚯蚓抵抗鎘毒性。鎘處理後,實驗中將有發生體壁凹陷的蚯蚓分成兩個部分,一為體壁凹陷部分,另一部分為沒有凹陷部分,體壁凹陷部位的鎘累積量遠高於沒有凹陷的部分,並且大多數體壁凹陷處都集中在蚯蚓身體的後半部,由此可知,體壁凹陷對於蚯蚓來說可能具有解毒的意義。除此之外,實驗中透過DNA 片斷化證明,體壁凹陷處其細胞有第一型細胞計畫性死亡(apoptosis)與細胞凋亡(necrosis)的發生,代表鎘處裡之下,會造成蚯蚓組織發生細胞死亡。鎘透過氧化壓力或者直接累積在組織而造成組織傷害,透過抗氧化劑的處理能有效的降低鎘毒性,並大幅提升存活率。而體壁凹陷的則是透過肌肉細胞死亡,造成組織崩解,導致外觀上的凹陷。 

Cadmium (Cd), a heavy metal commonly exists in environmental soil. Bioaccumulation of Cd causes systemic toxicity to most animals in the world. Earthworms have been recognized as suitable bio-indicator to evaluate soil pollution after exposure to heavy metals. In this study, Eisenia andrei was used as the animal model to study Cd-induced toxic response. Cd accumulation and mortality showed a dose-dependent and time-dependent in earthworms. After Cd exposure, earthworms showed several pathological phenomena including skin wrinkle, partial hyperemia, blood effusion, bleaching and abnormal body wall collapse. Cd-exposed earthworms showed higher H2O2 concentration and lower catalase activity. In addition, earthworms treated with various antioxidants (EUK134, curcumin, rosmarinic acid, catechin, or vitamin C) decreased both Cd accumulation and mortality. Furthermore, the collapse part of earthworms showed higher Cd accumulation than the non-collapse part in Cd-exposed earthworm, and body wall collapse usually occurred at the posterior part of earthworm. Nevertheless, cells underwent both apoptosis and necrosis at collapsed part. This may inferr that body wall collapse represent a detoxification mechanism in Cd-exposed earthworms.

摘要 1
Abstract 2
Introduction 3
1.1 Cadmium 3
1.1.1 The properties of cadmium 4
1.1.2 The industrial applications of cadmium 4
1.2 The cadmium toxicity to human and to other organisms 4
1.3 The role of oxidative stress in cell 5
1.3.2. Cadmium induced ROS 7
1.3.3. Antioxidants 7
1.4 Introduction of programmed cell death 9
1.4.3 Necrosis 11
1.5 Earthworm 11
1.5.1. Earthworm as a bio-indicator of polluted environment 12
1.5.2 Introduction of Eisenia Andrei 12
1.6 Purpose of study 12
Materials and Methods 13
2.1 Animal collection and maintenance 13
2.2 Cadmium exposure 13
2.3 Antioxidant treatment 13
2.4 Measurement of cadmium bioaccumulation 14
2.5 Tissue extract preparation 14
2.6 Protein concentration assay 14
2.7 H2O2 concentration 14
2.8 Catalase activity 15
2.9 Plastic embedded section and H&E stain 15
2.10 DNA fragmentation assay 16
2.11 Immunoblotting 16
2.12 Statistical Analysis 17
Results 18
3.1 Cd toxicity to earthworms, E. andrei 18
3.2 Muscles degenerated at body wall collapse part 18
3.3 Cd induces cell proliferation on epidermis and intestinal epithelium 19
3.4 Accumulation in earthworm E. andrei 20
3.5 Oxidative stress in the Cd-treated earthworms 21
3.6 Antioxidants can rescue Cd-exposed earthworm from Cd toxicity 21
3.7 The antioxidants pre-treatment prevented Cd accumulation in earthworms 22
3.8 Cd induce cell undergo apoptosis and necrosis 22
Discussion 23
4.1 Systemic Cd toxicity 23
4.2 Body wall collapse in Cd-exposed earthworm 23
4.3 Cd-induced oxidative stress to cause cellular damages 24
4.4 Possible mechanism of Cd-induced muscle degeneration 25
4.5 Biphasic role on Cd-induced cell proliferation 27
4.6 Cd-induced cell apoptosis 28
4.7 Rhabdomylysis 29
References 31
Figure 38

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