臺灣博碩士論文加值系統

　　鴨兒芹 (Cryptotaenia japonica Hassk) 是一種廣泛分佈於台灣中部山區的植物，可作為食用性蔬菜。本研究目的為探討鴨兒芹之乙醇萃取物 (ECJ) 在以高脂肪/高膽固醇飼料誘導之小鼠肥胖疾病動物模式下的降血脂作用。ICR小鼠隨機分成以下五組：(1) 餵食基礎飼料的正常飲食組；(2) 餵食高脂肪/高膽固醇飲食組；(3) 餵食高脂肪/高膽固醇飲食及20 mg/kg 的ECJ組；(4) 餵食高脂肪/高膽固醇飲食及100 mg/kg 的ECJ組；(5) 餵食高脂肪/高膽固醇飲食及simvastatin西藥治療組。實驗動物於8週後犧牲進行生化及組織病理檢驗。高脂肪/高膽固醇飲食合併餵食高或低劑量ECJ之小鼠的血清總膽固醇及三酸甘油酯分別比只餵食高脂肪/高膽固醇飲食組明顯地降低27.28% 及50.81%；肝臟之總膽固醇及三酸甘油酯含量測試也得到類似的趨勢。毒理檢查結果顯示在實驗劑量下並未發現餵食ECJ對實驗小鼠有任何肝腎毒性。此外，細胞學實驗結果發現ECJ在HepG2細胞中會抑制油酸 (oleic acid) 誘導的肝細胞內脂肪堆積。ECJ可減少HepG2細胞中參與三酸甘油酯和膽固醇合成相關分子的基因表現，並可增加腺苷酸單磷酸激酶 (adenosine monophosphate kinase, AMPK) 的磷酸化，顯示ECJ可能是透過AMPK的活化調控下游與脂質代謝相關分子的表現，進而減少肝細胞內的脂肪堆積。結論：本研究結果顯示鴨兒芹具有降血脂及抑制肝脂肪堆積之效用，可作為預防及減輕高血脂症的天然食材，建議民眾可於日常飲食中多加攝取。

　　Cryptotaenia japonica Hassk (mountain celery) is a common vegetable which is widely distributed in midland of Taiwan. The objective of this study was to investigate the hypolipidemic effects of Cryptotaenia japonica Hassk in a mouse obesity model induced by high fat/cholesterol diets (HFCD). ICR mice were fed with either HFCD or HFCD supplemented with 20 and 100 mg/kg Cryptotaenia japonica Hassk extract (ECJ, 95% ethanol extract) daily for 8 weeks and simvastatin as a therapeutic control. Plasma total cholesterol (TC) and triglyceride (TG) levels of mice combined fed with HFCD and ECJ were significantly reduced by about 27.28% and 50.81%, respectively, as compared to HFCD fed mice. Similar tendencies were observed in hepatic TC and TG. There were also no side effects observed on the functions of kidney and liver when feeding with various ECJ supplements. In cellular level, ECJ significantly reduced hepatic lipid accumulation in human HepG2 cells when stimulated with oleic acid. ECJ also downregulated the gene expressions of enzymes which involved in the cholesterol and triglyceride biosynthesis. We also found that ECJ increased the phosphorylation of AMPK in a dose-dependent manner. These results suggest that Cryptotaenia japonica Hassk prevented and mitigated high fat/cholesterol diet-induced hyperlipidemia as well as reduced hepatic lipid accumulation through AMPK activation pathway. Therefore, Cryptotaenia japonica Hassk could be used as a natural material against hyperlipidemia and hepatic steatosis.

縮寫表　i
摘要　iii
Abstract 　v
目次 　 vii
表次 　 xii
圖次 　 xiv
　
第一章　緒論　 1
1.1　研究動機 　1
1.2　研究架構　3
1.2.1　動物實驗　3
1.2.2　細胞實驗　4
　
第二章　文獻回顧　 5
2.1　高血脂與代謝性疾病　5
2.1.1　血漿脂質　5
2.1.2　脂蛋白　7
2.1.3　代謝症候群　9
2.2　脂肪肝疾病 　13
2.2.1　非酒精性脂肪性肝疾病定義 　14
2.2.2　非酒精性脂肪性肝炎與血脂之相關性　15
2.3　AMPK路徑　19
2.4　鴨兒芹文獻回顧 　21
　
第三章　材料與方法　23
3.1　儀器設備 　23
3.2　鴨兒芹萃取及成分檢測　23
3.2.1　萃取流程　23
3.2.2　HPLC分析條件　26
3.2.3　DPPH清除自由基實驗　26
3.3　動物實驗 (I) -降血脂療效評估 　 28
3.3.1　實驗動物飼養與分組　28
3.3.2　溶液配製與投予途徑　30
3.3.3　攝食量與體重記錄　30
3.3.4　動物犧牲、採血及臟器摘取　30
3.3.5　功能性指標評估　31
3.3.6　病理組織學切片與Haematoxylin & Eosin染色　31
3.3.7　肝臟中總脂質濃度測定 　 33
3.3.8　糞便脂質含量測定 　35
3.3.9　安全性指標評估　36
3.4　動物實驗 (II) -急毒性測試　36
3.4.1　實驗動物飼養與分組　36
3.4.2　溶液配製與投予途徑 　37
3.4.3　動物犧牲、採血及臟器摘取 　 38
3.4.4　血液學檢測 　38
3.4.5　血清生化檢測　38
3.4.6　臟器病理檢測 　39
3.5　細胞實驗 　39
3.5.1　細胞株來源及培養條件 　39
3.5.2　細胞毒性測試 　 39
3.5.3　脂肪油滴以Nile Red螢光染色定量 40
3.5.4　細胞內蛋白質之定性與定量分析 　40
3.5.5　反轉錄聚合酶鏈反應 　42
3.6　統計分析　45
　
第四章　結果　46
4.1　鴨兒芹萃取物成份分析　 46
4.1.1　HPLC分析結果　46
4.1.2　DPPH清除自由基實驗 　 49
4.2　動物實驗 (I) -降血脂療效評估 　55
4.2.1　功能性指標評估　55
4.2.2　安全性指標評估 　68
4.3　動物實驗 (II) -急毒性測試　72
4.3.1　臨床症狀及死亡率觀察 　72
4.3.2　血液學評估 　75
4.3.3　血清肝及腎臟功能指數評估 　77
4.3.4　臟器變化評估 　 79
4.4　細胞學實驗 　 84
4.4.1　細胞毒性測試 　 84
4.4.2　ECJ對HepG2細胞內脂肪堆積之影響 　85
4.4.3　ECJ抑制HepG2細胞內脂肪合成基因之表現　88
4.4.4　ECJ促進HepG2細胞內AMPK 之磷酸化　90
　
第五章　討論　92
5.1　動物實驗 　 92
5.2　細胞學實驗 　 97
　
第六章　結論　 100
　
參考文獻 　101
　
附圖 　116
附錄 　120

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