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標題: Apoptosis Induced by Ethanol Extract of Lactuca indica L. in HL-60 Cell Lines
作者: 陳伊萱
Chen, Yi-Hsuan
關鍵字: Lactuca indica L.;山萵苣;HL-60;Apoptosis;Cell cycle;HL-60細胞;程式性凋亡;細胞週期
出版社: 食品科學系
Lactuca indica L. belongs to the family Compositae, used as a folk medicine in anti-inflammatory, antibacterial, antidiabetic and other medications in Taiwan, China, Korea and Southeast Asia. The cancer occurrence is the leading cause of death for people in Taiwan; however, the research concerning the suppression tumor cells has not yet been investigated on this plant. The aim of this study was to evaluate the apoptosis inducing activity of ethanol, hot water, and cold water extracts of Lactuca indica L. on human leukemic HL-60 cell lines. Results showed that ethanol extracts of Lactuca indica L. exhibited a significantly apoptosis inducing activity in human leukemic HL-60 cells. The IC50 values for ethanol, hot water, and cold water extracts on apoptosis inducing activities was 0.31, 0.68, and >1 mg/mL, respectively. Flow cytometric analysis of the externalization of phosphatidylserine (PS) using the annexin V/PI method on ethanol extract of Lactuca indica L. treated HL-60 cells showed a concentration-dependent increase of apoptosis. Agarose gel electrophoresis of DNA from HL-60 cells treated with ethanol extract of Lactuca indica L. showed "ladder" pattern. Furthermore, ethanol extract of Lactuca indica L. can induce apoptosis in human leukemic HL-60 cells in a concentration- and time-dependant manner as determined by TUNEL positive and SubG1 population analyses. Therefore, the proliferation suppresses effect of ethanol extract of Lactuca indica L. on HL-60 cells may via a cell apoptosis mechanism. The ethanol extract of Lactuca indica L. contained quercetin, caffeic acid, rutin and chlorogenic acid of phenolic compound, and triterpenoid (330.4 mg/g ethanol extract of Lactuca indica L.). Cell viability and mitochondrial membrane potential (ΔΨ) collapses assay were performed on HL-60 cell line in comparison to four known compounds in Lactuca indica, quercetin, caffeic acid, rutin and chlorogenic acid. Cell viability showed that quercetin (IC50=50 μM) was the most growth inhibitory compound of Lactuca indica in the present study. Thus, quercetin might be the active compound in Lactuca indica L.

山萵苣(Lactuca indica L.)為菊科(Compositae)植物,在台灣、中國大陸及韓國等亞洲地區及東南亞地區均有分佈,傳統醫學記載其全草具有抗發炎、抑菌及降血糖等療效。癌症的發生至今仍位居國人十大死亡原因之首,目前並無山萵苣與抑制腫瘤細胞增生的相關研究。本研究擬選取山萵苣之冷水(CEL)、熱水(HEL)及乙醇(EEL)粗萃取物,探討其對HL-60細胞增生之抑制效應。結果顯示三種萃取物中以EEL抑制效果最佳,IC50為0.31 mg/mL,其次依序為HWL (0.68 mg/mL)>CEL (>1 mg/mL)。HL-60細胞以EEL處理48小時,其細胞膜之磷脂質絲胺酸(PS)有顯著外翻的現象,細胞膜電位亦有降低之情形。DNA電泳分析發現,EEL可顯著誘發DNA 片段化,且樣品添加劑量呈現正相關,並與TUNEL assay有一致性的結果。以流式細胞儀分析HL-60細胞週期變化,結果發現以EEL (1 mg/mL)處理48小時,其sub-G1 phase可顯著提高至42.5%,呈現G1 phase arrest效應,因此EEL對於抑制HL-60腫瘤細胞增生之效應可能與誘發細胞凋亡(apoptosis)有關。成分分析研究發現,EEL萃取層含有Quercetin、Caffeic acid、Rutin、Chlorogenic acid等酚類化合物及330.4 mg/g EEL的三萜類化合物;利用山萵苣中所含Quercetin、Caffeic acid、Rutin及Chlorogenic acid處理HL-60細胞,結果發現Quercetin抑制細胞存活之能力最佳,IC50濃度約為50 μM,此結果與粒線體膜電位的降低情形相似。綜合實驗結果推測,山萵苣乙醇萃取物確實具有誘發細胞凋亡之能力,Quercetin可能為其主要活性成分之一。
Appears in Collections:食品暨應用生物科技學系

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