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    Please use this identifier to cite or link to this item: http://asiair.asia.edu.tw/ir/handle/310904400/26167


    Title: 台灣中草藥紫背草之檢測及肝毒性之研究
    Authors: 楊雅甄
    Contributors: 健康學院;保健營養生技學系
    Keywords: 紫背草;吡咯啶類生物;氣相層析質譜儀和流式細胞儀;Emilia Sonchifolia (L);DC;Pyrrolizidine Alkaloids;flow cytometry and GC/MS
    Date: 2012
    Issue Date: 2013-07-18 15:36:13 (UTC+8)
    Abstract: 紫背草(Emilia sonchifolia (L.) DC.) 別名葉下紅、紅背草、一點紅、羊蹄草,在中草 藥中有清火、消炎解毒、解熱退燒等功效。本研究擬收集全台各地之紫背草,以 0.5 M 鹽酸 溶液萃取,陽離子交換固相萃取濃縮,經氣相層析質譜儀(GC/MS)分析檢測紫背草所含肝毒性 及致癌之吡咯啶類生物鹼 (pyrrolizidine alkaloids;PAs)。目前已檢測出紫背草中含有二種類型 的PAs:(1) retronecine 類型,即senecionine 和 integerrimine;(2) otonecine 類型,即senkirkine、 neosenkirkine 和acetyl senkirkine 等化合物,但尚有五個PAs 結構鑑定中。本研究定量擬以methyl stearate 為內標準品,以senkirkine 為標準品製作檢量線定量各PAs 的含量,檢量線濃度擬為範 圍20-400 μg/ml。 最 近 其 致 毒 機 制 研 究 顯 示 吡 咯 烷 士 啶生物鹼之致腫瘤性, 係經由形成 6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizidine (DHP) 衍生物之DNA 加合物導致基因 毒性機制。然而,迄今PAs 對抗人類肝細胞的作用機轉之研究尚嫌不足。本計畫對PAs 誘導 肝細胞計畫性死亡的研究如下: (一) 利用 MTT 與trypan blue 染色實驗,探討PAs 對肝細胞株的細胞毒性作用 (二) 利用 TUNEL 的DNA 損害實驗評估PAs 對肝細胞株DNA 細胞計劃性死亡的情形。 (三) 利用活性氧物質的探針與流式細胞儀的技術,我們將探討PAs 是否會大量誘導肝細胞內 源性活性氧物質的產生。 (四) 利用流式細胞儀的技術,我們將探討 PAs 是否會影響肝細胞內GSH 的耗空。 (五) 利用抗氧化酵素活性分析實驗,我們將探討 PAs 是否會影響肝細胞內抗氧化酵素,例如 超氧歧化酵素(SOD)與過氧化酵素(catalase)的活性。

    Emilia sonchifolia (L.) DC. (Lilac tasselflower, Yi dian hong, Yang ti cao) is used there in folk medicine as antipyretic, as a remedy against influenza. We have investigated five compounds of the hepatotoxic and tumorigenic pyrrolizidine alkaloids (PAs) contained in Emilia sonchifolia (L.) DC. in Taiwan using 0.5 M HCl(aq). Two structural types of PAs were extracted and identified from the dried plant material by cation-exchange solid-phase extraction (CE-SPE) and GC/MS method: (1) otonecine bases, e.g. senkirkine, acetyl senkirkine and neosenkirkine; (2) retronecine bases, e.g. senecionine and integerrimine. There are five other compounds under identifying. Quantitation of the individual PAs will be determined by linear regression curve in the range of 20-400 μg/mL of senkirkine and methyl stearate as an internal standard examined. Recent mechanistic studies indicate that pyrrolizidine alkaloids induce tumors via a genotoxic mechanism mediated by 6,7-dihydro-7-hydroxy-1-hydroxymethyl- 5H-pyrrolizidine (DHP) DNA derived adducts formation. However, the effects of PAs-induced apoptosis on human liver chang’s cell have not been well studied. In order to evaluate the importance of these events in PAs-induced apoptosis, we propose to study the following: (1) Using MTT and trypan blue exclusion assay, we want to study the cytotoxicity of human liver chang’s cell lines. (2) Using TUNEL assay, we want to investigate the DNA fragmentation during PAs inducing apoptosis in human liver chang’s cell. (3) Using the probes of ROS and flow cytometric technique, we want to confirm the increasing intracellular ROS during PAs inducing human liver chang’s cell death. (4) Using flow cytometric technique, we want to evaluate the changing of intracellular GSH during PAs inducing human liver chang’s cell death. (5) Using the antioxidant enzyme assays, we want to assay the activity of antioxidant enzymes, such as superoxide dismutase and catalase during PAs inducing apoptosis.
    Appears in Collections:[食品營養與保健生技學系] 科技部研究計畫

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