本研究建立海鱺前脂肪細胞培養系統，並將此系統應用於探討主要飼料脂質或脂肪酸對海鱺脂肪細胞增殖和分化程度、脂肪蓄積量及脂肪分化相關基因表現的影響，期能應用本系統結果，透過飼料脂肪或脂肪酸配方，調控海鱺魚體脂肪含量及分布。海鱺為少數可以餵飼高脂 (>17%) 飼料之溫水性海水魚種，且市場歡迎其肌肉具高油脂含量，能夠瞭解飼糧因子對脂肪細胞增殖及脂肪細胞油脂蓄積的調控，對海鱺養殖產出符合市場需求高質產品將有很大益處。研究分成四實驗， 包括 (實驗一)前脂肪細胞培養系統建立、(實驗二) 飼料脂肪酸在飼育實驗 (in vivo)與細胞實驗 (in vitro) 基因表現和脂質累積的相對應結果、(實驗三)單一脂肪酸與(實驗四)複合脂肪酸對前脂肪細胞分化基因 CCAAT/enhancer binding protein- β (C/EBPβ)、peroxisome proliferator-activated receptors (PPARs)、fatty acid binding protein1 (FABP1)、fatty acid transport protein 2 (FATP2)、和fatty acid synthase (FAS) 及細胞脂質累積的影響。
實驗一結果顯示海鱺前脂肪細胞最適培養條件為10% 胎牛血清的L-15培養液在28 °C恆溫培養，18天可增生至90% 滿盤 (confluence)。經insulin-transferrin-selenium誘導初期分化後，於培養液添加10%脂質混合物 (lipid mixture)維持細胞分化，分化20天為成熟脂肪細胞，進行Oil Red O染色測定脂質累積量與即時定量PCR (real-time PCR)分析發現，成脂基因(adipogenic genes) FABP1、FATP2和FAS表現量會隨著分化時間而增加，於成熟脂肪細胞達到最高值。
實驗二比較飼育實驗與細胞實驗在評估脂肪生成之研究之一致姓，使用相同脂肪酸組成處理時，含n-6脂肪酸為主的紅花油、橄欖油和棕櫚油飼料脂肪酸組，不論是飼育實驗和細胞實驗均比含n-3脂肪酸為主的魚油和紫蘇油組更能增加脂質累積的效果，且in vitro與in vivo實驗PPARs和FABP基因表現量均呈現正相關，顯示海鱺前脂肪細胞實驗結果與海鱺飼育實驗脂肪分析結果相似度高。
實驗三探討單一脂肪酸棕櫚酸 (palmitic acid，PA 16:0)、硬脂酸(stearic acid，SA 18:1 n-9)、油酸 (oleic acid，OA 18:1 -9)、亞麻油酸(linoleic acid，LA 18:2 n-6)、次亞麻油酸(α-linolenic acid，LNA 18:3 n-3)、Eicosapentaenoic acid (EPA 20:5 n-3)或Docosahexaenoic acid (DHA 22:6 n-3)七種脂肪酸對海鱺前脂肪細胞分化轉錄因子、成脂基因與脂質累積各有不同的影響。轉錄因子C/EBPβ和PPARs在分化初期即表現，且C/EBPβ和PPARγ基因表現時間比PPARα和PPARβ略早，成脂基因FABP1、FATP2和FAS則在分化中末期才開始表現。EPA、DHA、OA和LA使C/EBPβ和PPARs顯著表現，OA和EPA顯著增加FABP1、FATP2和FAS基因表現量。OA顯著增加脂肪細胞尺寸與脂肪細胞累積脂質量，而DHA抑制脂肪細胞肥大，且LNA和DHA明顯不利於油脂累積。
棕櫚油、芥花籽油、大豆油、亞麻仁油、鯷魚油和鱈魚肝油是魚類飼料主要使用的油脂，實驗四將這些油脂的主要脂肪酸以複合方式加入培養液，瞭解多個脂肪酸的協同或拮抗效應。複合油脂中含有OA、LA、EPA和DHA時，均會增加轉錄因子 (C/EBPβ和PPARγ) 基因表現，此四個脂肪酸彼此有協同作用但只有OA與濃度有關；一旦含有PA、SA或LNA，會產生拮抗作用而降低轉錄因子基因表現，其中LNA僅只要存在少量即會產生抑制效果。PA、OA和LA存在顯著提升FAS基因表現和三酸甘油酯 (TG)累積，且三個脂肪酸之間有協同作用；LNA、EPA和DHA均會降低FAS表現和TG含量。LNA、EPA和DHA存在會使脂肪細胞產生較高的活性氧物種，增加細胞氧化壓力。
本研究結果綜合顯示，富含LA與OA的油脂有助於增加海鱺脂肪細胞油脂蓄積，而含LNA的油脂不利於脂質累積，應用海鱺前脂肪細胞培養系統來推測營養素對海鱺脂肪生成的影響似無窒礙之處。

The present study established cobia preadipocytes culture system, and used the system to investigate the effects of dietary lipids or fatty acids on cobia preadipocytes proliferation, differentiation, lipid accumulation and adipogenesis genes expression. The aim was to explore the possibility modulate cobia lipid distribution and content through nutritional manipulation. Cobia is a warmwater finfish that accepts high lipid (>17%) diets and is well valued for its white muscle of high lipid levels. In the present study, in addition to optimizing the culture medium for the differentiation of preadipocytes model in Experiment I, several studies evaluating the effects of fatty acids were carried out, including: (Experiment II) comparisons of lipid accumulation and differentiation genes expression in adipocytes between in vivo and in vitro experimental settings, (Experiment III) the effects of single fatty acid as well as (Experiment IV) multiple fatty acids on cobia preadipocytes differentiation genes CCAAT/enhancer binding protein- β (C/EBPβ), peroxisome proliferator-activated receptors (PPARs), fatty acid binding protein1 (FABP1), fatty acid transport protein 2 (FATP2) and fatty acid synthase (FAS) expression.
In the Experiment I, cobia preadipocytes cultured in L-15 medium with 10% FBS at 28 °C was found to have a high proliferation potential leading to a monolayer confluence by day 18. Oil Red O staining indicated numerous large cytoplasmic lipid droplets were formed and real-time PCR analyses showed increasing upregulations of FABP1, FATP2 and FAS as preadipocytes differentiated.
In the Experiment II, the expression levels of PPARs and FABP were compared between an in vivo and an in vitro experiments. The results of the in vitro and in vivo experiments were found to be compatible. Lipid accumulation was found to be greater in response to safflower oil, olive oil and palm oil treatments in both experiments than in fish oil and perilla oil treatments. At both molecular and cell levels, the effects of fatty acids on adipogenesis in vitro and in vivo systems showed similar trends.
In the Experiment III, palmitic acid (PA 16:0), stearic acid (SA 18:1 n-9), oleic acid (OA 18:1 -9), linoleic acid (LA 18:2 n-6), α-linolenic acid (LNA 18:3 n-3), eicosapentaenoic acid (EPA 20:5 n-3) or docosahexaenoic acid (DHA 22:6 n-3) were found to modulate differentially on cobia preadipocyte differentiation. C/EBPβ and PPARs expression increased during the early phase of differentiation and decreased at later stages of differentiation. On the other hand expressions of FABP1, FATP2 and FAS were increased throughout the differentiation process. The expression levels of C/EBPβ and PPARγ increased when the cells were exposed to OA, LA, EPA and DHA groups. OA and EPA increased the expression levels of FABP1, FATP2 and FAS. LNA and DHA led to a lower accumulation of triglycerides (TG) in mature adipocytes than OA.
In the Experiment IV, the synergistic or antagonistic effects of major fatty acids of palm oil, rapeseed oil, soybean oil, flaxseed oil, anchovy oil or cod liver oil on adipogenesis and adipocyte differentiation were investigated. The expression levels of C/EBPβ and PPARγ were increased when the adipocytes were exposed to OA, LA, EPA or DHA, indicating a synergistic effect. Transcription factor genes expression levels were reduced with the exposure to PA, SA and low levels of LNA. The expression level of FAS and TG accumulation were significantly higher for PA, OA and LA than for LNA, EPA and DHA. The amount of reactive oxygen species production increased in adipocytes with exposure to LNA, EPA and DHA.

論文審定書∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ i
誌謝 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ ii
中文摘要 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ iii
英文摘要 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ vi
目錄 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ ix
圖次 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ xi
表次 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ xiv
英文縮寫對照表∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ xv
第一章 文獻回顧∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 1
1.1 水產飼料中的油脂∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 1
1.2 魚油取代對體組成及飼料效率的影響∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 3
1.3 脂肪形成過程∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 9
1.4 脂肪細胞分化標誌∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 11
1.5 多元不飽和脂肪酸∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 32
1.6 細胞實驗使用脂肪酸的研究∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 41
第二章 前言∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 44
第三章 綜合材料與方法∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 51
3.1 海鱺前脂肪細胞分離方式∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 51
3.2 細胞培養成長與分化條件∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 51
3.3 前脂肪細胞增生抗原測定∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 52
3.4 前脂肪細胞分化為脂肪細胞程度之測定∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 52
3.5 脂肪細胞Total RNA 萃取∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 53
3.6 RNA 變性電泳分析∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 54
3.7 互補去氧核醣核酸(cDNA)之製備∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 54
3.8 PCR 及DNA 電泳分析∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 54
3.9 即時定量PCR∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 55
3.10 統計分析∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 55
第四章 建立海鱺前脂肪細胞培養系統 (實驗一) ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 58
4.1 前言∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 58
4.2 材料與方法∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 59
4.3 結果∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 61
4.4 討論∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 68
第五章 脂肪酸影響海鱺前脂肪細胞脂肪分化基因表現與脂質累
積：細胞培養與飼養試驗之比較(實驗二) ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙
71
5.1 前言∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 71
x
5.2 材料與方法∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 72
5.3 結果∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 76
5.4 討論∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 80
第六章 單一脂肪酸對海鱺前脂肪細胞分化及基因表現之影響
(實驗三) ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙
88
6.1 前言∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 88
6.2 材料與方法∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 89
6.3 結果∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 90
6.4 討論∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 98
第七章 複合脂肪酸對海鱺前脂肪細胞分化及油脂累積之影響
(實驗四) ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙
107
7.1 前言∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 107
7.2 材料與方法∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 110
7.3 結果∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 114
7.4 討論∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 123
第八章 總結論∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 133
第九章 參考文獻∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 135
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