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標題: | 金針花乙醇萃取物可藉由提升小鼠肝中抗氧化酵素活性及降低發炎反應以減緩酒精性肝損傷 Liver Protective Effect and Mechanism of Ethanol Extract of Daylily Flower against Alcoholic Liver Damage in Mice by Enhancing Activity of Antioxidative Enzymes and Reducing Inflammation Responses |
作者: | Ying-Chu Chen 陳映竹 |
指導教授: | 沈立言(Lee-Yan Sheen) |
關鍵字: | 酒精性肝病,金針花,氧化壓力,抗氧化,NF-E2-related factor 2, alcoholic liver disease,daylily flower,oxidative stress,antioxidant capacity,NF-E2-related factor 2, |
出版年 : | 2014 |
學位: | 碩士 |
摘要: | 肝病之病程常伴隨著脂肪浸潤、脂肪肝、肝發炎等病狀,最終有可能惡化成肝纖維化、肝硬化甚至肝癌。有許多原因會導致肝病的發生,當飲酒過度時,體內大量代謝酒精所產生的氧化壓力,會引起酒精性肝病(alcoholic liver disease, ALD)之形成。金針花(Hemerocallis fulva L.)為台灣著名農產品之一,為中國傳統食療方,近年研究發現,金針花含有天然抗氧化物質,如類胡蘿蔔素、類黃酮及花青素等。此外,金針花萃取物已被證實在脂多醣(lipopolysaccharide, LPS)誘導巨噬細胞中可抑制nitric oxide(NO)的產生及降低誘導型一氧化氮合酶(inducible nitric oxide synthase, iNOS)表現,且具有清除活性氧能力。有研究指出,金針花經以95%之乙醇萃取後,與其水萃物相較之下,具有較佳之抗氧化活性,與其富含多酚類物質有關,其中又以芸香素(rutin)之含量為最高。因此,本研究的假說認為金針花乙醇萃取物因富含抗氧化活性而具有減緩酒精所造成肝損傷的效果。本研究以Lieber-DeCarli酒精液態飼料誘導小鼠酒精性肝損傷之動物模式,進行金針花乙醇萃取物之護肝功效評估。實驗動物分成五組,包括正常控制組(Control)、酒精誘導肝損傷負對照組(ALD)及酒精誘導肝損傷並同時給予金針花乙醇萃取物低劑量(400 mg/kg bw/day, ALD + LDFE)、高劑量(800 mg/kg bw/day, ALD + HDFE)及金針花乙醇萃取物中純物質-芸香素(2.6 mg/kg bw/day, ALD + Rutin)處理組。實驗結果顯示,相較於ALD組,餵食金針花乙醇萃取物低劑量及高劑量組,其血清中肝臟生化功能指標酵素天門冬胺酸轉胺酶(aspartate transaminase, AST)及丙胺酸轉胺酶(alanine transaminase, ALT)數值具有顯著降低的現象;肝臟組織病理學觀察方面,餵食低劑量及高劑量金針花乙醇萃取物,可改善肝臟中脂肪堆積之情形。此外,金針花乙醇萃取物亦可提高肝臟中抗氧化物質麩胱甘肽(glutathione, GSH)之含量,及提升肝臟中抗氧化酵素之活性,同時改善小鼠肝臟中脂質過氧化及發炎反應之情況,且向下調控肝中細胞激素P450 2E1(cytochrome P450 2E1, CYP2E1)酵素之活性。進一步以西方墨點法及免疫組織染色法,評估金針花乙醇萃取物之抗氧化機制,結果顯示金針花乙醇萃取物可提升NF-E2-related factor 2(Nrf2)於細胞核之表現量及其下游蛋白heme oxygenase-1(HO-1)表現量。綜合以上實驗結果,推測金針花乙醇萃取物可能藉由其抗氧化和抗發炎作用,以及向下調控CYP2E1之表現量,並活化Nrf2/HO-1,而具有改善酒精性肝損傷之護肝功效。 Liver disease is one of the most widely contracted diseases in Taiwan. There are many reasons result in liver disease, one of those are excessive alcohol consumption which will lead to alcoholic liver disease (ALD), such as in fatty liver, alcoholic hepatitis and end to cirrhosis, which results to hepatocyte dysfunction. The pathogenesis of ALD is related to lipid accumlation, oxidative stress, and inflammation. The flower of daylily (Hemerocallis fulva L.) is widely used in traditional medication and food material in eastern Asia. Previous studies indicated that the flowers of daylily are rich in variety of antioxidants such as carotenoids, flavonoids, anthocyanin, and so on. Furthermore, the extracts of daylily flowers have been reported to have the inhibition efficiency on the nitric oxide (NO) production, reducing the inducible nitric oxide synthase (iNOS) induction in lipopolysaccharide (LPS)-activated macrophages; and reative oxygen species (ROS) scavenging activity. Recent study found that the 95% ethanol extracts of daylily flowers content high level of polyphenols and exhibited better antioxidant activities than water extracts. The hypothesis of this study is ethanol extracts of daylily flower (DFE) may have the hepatoprotective effects on alcoholic liver disease mice by ameliorating the hepatic oxidative stress and inflammatory responses.. The aim of this study is to investigate the liver protective capability of ethanol extract of daylily flowers in male C57BL/6 mice which feed with Lieber-DeCarli alcohol-containing liquid diet, and its hepatoprotective mechanism. Mice were divided five groups: fed with normal liquid diet (normal control group), ethanol-containing liquid diet is (negative control group, ALD) or ethanol-containing liquid diet treated with low dosage of ethanol extract of daylily flower (DFE) at 400 mg/kg bw/day (ALD + LDFE group), high dosage of DFE at 800 mg/kg bw/day (ALD + HDFE group), and rutin which content as high dosage of DFE at 2.6 mg/kg bw/day (ALD + Rutin group) for 4 weeks. Our results showed that in comparison with the control group, the ALD group showed liver injury as evidenced by histological changes and elevation in serum biochemical, liver inflammation, and oxidative stress. These pathophysiological changes were attenuated by DFE supplementation. Further studies demonstrated an inhibitory effect of DFE on the critical role in ALD, cytochrome P450 2E1 (CYP2E1). DFE also attenuated alcohol-induced reduction in heme oxygenase-1 (HO-1) and NF-E2-related factor 2 (Nrf2). According to our results, the hepatoprotective effects of DFE were shown to be associated with antioxidative and anti-inflammatory effects as well as the downregulation expression of CYP2E1, and enhancement of HO-1 and Nrf2 expression in liver. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57069 |
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顯示於系所單位: | 食品科技研究所 |
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