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研究生:黃福龍
研究生(外文):Fu-Long Huang
論文名稱:台灣產桂花之安全性、抗致敏性氣喘及護肝功能之評估
論文名稱(外文):Valuation on the safety and anti-allergy asthma and hepatoprotection of Osmanthus fragrans flower in Taiwan
指導教授:洪敏元 洪千雅
指導教授(外文):Min-Yuan Hung Chien-Ya Hung
學位類別:碩士
校院名稱:中華醫事學院
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:94
語文別:中文
論文頁數:136
中文關鍵詞:桂花氣喘護肝
相關次數:
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本研究主要探討台灣產桂花的安全性以及抗致敏性氣喘與抑制 acetaminophen 對肝細胞損傷的效果。
第二章是桂花萃取物安全性評估,分為急性毒性試驗及慢性毒性試驗,急性毒性試驗為 BALB/c 小鼠經管灌分別餵食 5.0 g/kg B.W. (5 g KFE) 及 10.0 g/kg B.W. (10 g KFE) 75 % 乙醇桂花萃取物,觀察72小時。實驗結果顯示各實驗組小鼠均無死亡之現象。慢性毒性試驗為 BALB/c 小鼠連續 28 天經管灌分別餵食低劑量 100 mg/kg B.W. (LEKF)、中劑量 800 mg/kg B.W. (MEKF)、高劑量 1000 mg/kg B.W. (HEKF) 75 % 乙醇桂花萃取物及低劑量 10 mg/kg B.W. (LR)、高劑量 100 mg/kg B.W. (HR) 抗氧化城分 rutin。實驗結果顯示各實驗組均無死亡之現象,且無外觀上之中毒現象產生。與控制組相比較體重及相對器官重量均無顯著性差異。肝臟及腎臟無脂質過氧化現象產生。於肝、脾、肺、腎臟組織病理切片中,發現無特殊之病變發生。綜合以上結果顯示,桂花萃取物及其抗氧化成分 rutin 經管灌餵食 BALB/c 小鼠連續 28 天,不會造成小鼠中毒之現象,證實桂花萃取物是無毒性的物質。
第三章是桂花萃取物抗氣喘能力研究。小鼠經抗原誘導產生氣喘時也會造成肺、肝臟氧化現象產生 TBARs 值上升。連續 28 天管灌餵食不同劑量 75 % 乙醇桂花萃取物及不同劑量抗氧化成分 rutin,可以改善經 OVA 抗原刺激誘導 BALB/c 小鼠氣喘症狀的能力。實驗結果顯示各實驗組 [ 桂花萃取物組: 100 mg/kg B.W. + OVA (LEKF)、800 mg/kg B.W. + OVA (MEKF)、1000 mg/kg B.W. + OVA (HEKF);rutin 組: 10 mg/kg B.W. + OVA (LR)、100 mg/kg B.W. + OVA (HR) ] 與控制組 [Positive control (PS)、Negative control (NS) ] 相比較,體重及相對器官重量均無顯著差異。於血清中檢測抗體發現,實驗組均能明顯降低氣喘發生時 IgE 抗體的產生、Eosinophil 發炎細胞的增生及呼吸道阻力現象,以及肺細胞與肝細胞脂質氧化現象,並從肺臟組織病理切片顯示,能降低氣喘小鼠支氣管黏膜發炎細胞及分泌物沉積。使氣喘小鼠有偏走向 Th1 分泌途徑,而不完全走向導致氣喘之 Th2 分泌途徑。
第四章是桂花萃取物護肝能力研究,BALB/c 小鼠管灌餵食 75 % 乙醇桂花萃取物 (LEKF、MEKF、HEKF) 及抗氧化成分 rutin (LR、HR),連續15天,於小鼠犧牲前三小時腹腔注射 acetaminophen (1.0 g/kg B.W. ),實驗結果顯示,各實驗組能明顯降低 acetaminophen 所導致 GOT 值的上升,減少肝臟及肺臟脂質過氧化現象。於組織病理切片檢驗中,實驗組均能明顯降低 acetaminophen 所造成小鼠肝臟組織空泡化之傷害現象。綜合以上結果顯示,桂花及其抗氧化成分 rutin 經管灌餵食 BALB/c 小鼠連續 15 天,確實能夠降低 acetaminophen 藥物毒性傷害,達到保護肝細胞的能力。
The aim of this study was to investigate the safety, anti-allergy asthma, and hepatic protection of Osmanthus fragrans flower in Taiwan on acetaminophen-induced hepatic damage in BALB/c mice.
Chapter 2 was evaluates the acute and the 28 days subacute toxicities of O. fragrans flower extract (Kwei-Fah extract; KFE) by 75 % ethanol. In the acute toxicity study, BALB/c mice were single orally administered with 5.0 g/kg (5g KFE) and 10.0 g/kg B.W. (10g KFE) dosages of Kwei-Fah extract to obtain the LD50 value under 72 hr. According to the definition of acute toxicity, Kwei-Fah extract was practically nontoxic (LD50: >10 g/kg BW). In 28 days subacute toxicity study, the body and visceral organ weights of BALB/c mice administered with 100 mg to 1000 mg/kg B.W. dosages of KFE and its active compounds rutin (10 and 100 mg/kg B.W. mice) did not apparently change. The histopathology of visceral organ and the values of GOT, GPT and BUN in serum did not significantly different from those of the control group. There were no significant changes in lipid peroxidation in liver and kidney. The results suggested that the KFE and rutin had no obvious toxicity on BALB/c mice.
Chapter 3 was evaluates the anti-allergic asthma of Kwei-Fah extract by 75 % ethanol in BALB/c mice. Ovalbumin (OVA)-induced asthma of BALB/c mice had significantly increased lung and liver TBARS value. The oral treatment with different dose of KFE (100, 800 and 1000 mg / kg B.W. mice) and rutin (10 and 100 mg/kg B. W. mice) had significantly reduced OVA-specific IgE in serum, the eosinophils percentage of total leukocytes from bronchoalveolar lavage fluid (BALF), methacholine-induced airway hyperresponsiveness (AHR), and decreased lipid peroxidation in lung and liver cell of BALB/c mice. Therefore, the results suggest that KFE and rutin may decrease the Th2-related allergic disease on BALB/c mice.
Chapter 4 was evaluates the hepatic protection of 75 % ethanol Kwei-Fah extract on acetaminophen-induced hepatic damage in BALB/c mice. BALB/c mice were single orally administered with different dosages of KFE (100, 800 and 1000 mg / kg B.W. mice) and rutin (10 and 100 mg/kg B. W. mice) for 15 days, and then injected with 1.0 g acetaminophen /kg B.W. (APAP). The result showed that KFE and rutin significantly decreased the GOT level in serum and lipid peroxidation in liver and lung cell. Liver histopathology showed that KEF and rutin reduced the incidence of liver lesions including cytoplasmic vacuolization. The results indicated that KFE and rutin could protect liver against acetaminophen-induced damage.
總目錄

圖 次...........................................................................................................v
表 次.........................................................................................................vii
中文摘要....................................................................................................viii
英文摘要.......................................................................................................x
Abbreviation................................................................................................xii


第一章 文獻整理
一、 自由基與抗氧化.....................................................................1
二、 過敏性氣喘.............................................................................3
(一) 氣喘定義..........................................................................4
(二) 氣喘反應機轉..................................................................4
(三) 氣喘成因與症狀..............................................................5
(四) 氣喘與氧化壓力………………………………….….…6
(五) 氣喘治療………………………………………………..6
三、 Acetaminophen (APAP)…………………………….….........7
(一) Acetaminophen 毒性機制……………………….…......7
(二) Acetaminophen 毒性與氧化壓力………………….......8
四、 桂花相關之研究……………………………………………10
五、 研究目的…………………………………………………....13
六、 研究架構…………………………………………………....14


第二章 桂花萃取物及 rutin 安全性評估
一、 摘 要……………………………………………………....16
二、 材料與方法………………………………………………....17
(一) 桂花 75 % 乙醇萃取物製備………………………….17
(二) 動物實驗設計(急性毒性試驗、慢性毒性試驗)……...17
(三) 動物犧牲及實驗檢體收集與分析…………………….21
(四) 統計分析…………………………………………..…...25
三、 結果與討論……………………………………………..…..26
(一) 實驗劑量之選擇………………………………….…....26
(二) 臨床症狀……………………………………………….27
(三) 試驗期間小鼠體重變化……………………………….27
(四) 臟器重量變化…………………………………………28
(五) 血液分析………………………………………………28
(六) 肺沖洗液血球變化……………………………………29
(七) 臟器組織病理檢查……………………………………30
(八) 臟器脂質過氧化………………………………………30
四、 結 論………………………………………………………31


第三章 桂花萃取物及 rutin 抗致敏氣喘功能之評估
一、 摘 要………………………………………………………46
二、 材料與方法…………………………………………………47
(一) 桂花 75 % 乙醇萃取物製備………………………….47
(二) 動物實驗設計………………………………………….47
(三) 動物犧牲及實驗檢體收集與分析…………………….50
(四) 統計分析…………………………………………….…58
三、 結果與討論………………………………………………….59
(一) 試驗期間小鼠體重變化…………………………….….59
(二) 臟器重量變化…………………………………………..59
(三) OVA 特異性抗體 IgE、IgG1、IgG2a 分析………….59
(四) 肺沖洗液血球變化…………………………………......60
(五) 呼吸道阻力………………………………………….….60
(六) 臟器組織病理檢查……………………………………..61
(七) 臟器脂質過氧化………………………………………..61
四、 結 論……………………………………………………….63


第四章 桂花萃取物及 rutin 護肝功能之評估
一、 摘 要………………………………………………………79
二、 材料與方法…………………………………………………80
(一) 桂花 75 % 乙醇萃取物製備………………………….80
(二) 動物實驗設計………………………………………….80
(三) 動物犧牲及實驗檢體收集與分析…………………….82
(四) 統計分析……………………………………………….85
三、 結果與討論………………………………………………....86
(一) 試驗期間小鼠體重變化……………………………….86
(二) 臟器重量變化………………………………………….86
(三) 血液分析……………………………………………….86
(四) 臟器組織病理檢查…………………………………….87
(五) 臟器脂質過氧化……………………………………….87
四、 結 論………………………………………………………88

第五章 參考文獻………………………………………………………101


附 錄 (已發表之桂花相關研究)
一、桂花萃取物對 acetaminophen 誘導 BALB/c 小鼠肝毒性影響之
研究…………………………………………………………………111
二、桂花市售製品抗氧化能力、總多酚含量及抗氧化指標成分verbascoside 及 rutin 之含量分析……………………………...124
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