以初代細胞探討辣木葉萃取物之免疫調節作用

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Title:	以初代細胞探討辣木葉萃取物之免疫調節作用 Immunomodulatory effects of Moringa oleifera leaves extracts in murine primary culture
Authors:	吳冠穎
Contributors:	保健營養系夏彩蘭
Keywords:	脾臟腸繫膜淋巴結皮耶氏斑辣木葉細胞激素小鼠 spleen mesenteric lymph nodes Peyer’s patches Moringa oleifera cytokine mice
Date:	2013
Issue Date:	2014-03-11 14:22:28 (UTC+8)
Abstract:	辣木原產於印度，因果莢似鼓槌，又稱為鼓鎚樹（Drumstick），是多年生熱帶落葉喬木，全世界已知有14個品種。辣木因為容易繁殖、生長快速，又具有豐富的營養素，所以在亞洲辣木葉、花和未成熟的種子普遍被當作食用蔬菜和民間草藥使用。近來也有越來越多的文獻指出辣木葉萃取物、種子或植株對免疫細胞株具有抗腫瘤或抗發炎的功效，但沒有對初代免疫細胞的相關研究文獻。因此本研究的目的是探討辣木葉萃取物對於小鼠脾臟、皮耶氏斑和腸繫膜淋巴細胞分泌Th1/Th2細胞激素之影響。8-10週齡的BALB/c雌鼠，其脾臟、皮耶氏斑和腸繫膜淋巴結製成單一細胞懸浮液，分別加入不含或含 10、50、100、200、500及1000μg/mL濃度的辣木葉水、乙醇和乙酸乙酯萃取物、10 μg/mL LPS和10 μg/mL Con A共同培養24和48小時後，取上清液測IL-2、IL-4、IL-6、IL-10、IL-12p70及IFN-γ的含量。以不含辣木葉萃取物作為對照組。以MTT試驗測試辣木葉萃取物對細胞之毒性。冷凍乾燥辣木葉水、乙醇和乙酸乙酯萃取物回收率分別為22.16±0.59 %、 23.07±1.09 % 和10.47±0.46 %。熱風乾燥辣木葉沸水、乙醇和乙酸乙酯萃取物回收率分別為 38.1± 3.96%、24.24 ± 0.08 %和7.58 ± 2.09 %。冷凍乾燥辣木葉水和乙醇萃取物刺激小鼠脾臟、MLN和PP分泌細胞激素的效果比熱風乾燥辣木葉佳。辣木葉乙酸乙酯萃取物不影響脾臟、MLN和PP細胞分泌細胞激素。本研究結果顯示，辣木葉萃取物刺激IL-10的分泌較其他細胞激素顯著，由此推論辣木葉萃取物可能藉由IL-10分泌增加而促進體液性免疫反應。關鍵字：脾臟、腸繫膜淋巴結、皮耶氏斑、辣木葉、細胞激素、小鼠 Moringa oleifera is native to India and is also known as the 'drumstick' tree. There are 14 known species of the genus Moringa, which grow in the tropics. The young leaves, flowers and immature pods of drumstick tree are a common edible vegetable in Asian diets because it is easy to breed, grows quickly and contains nutritive value. Some studies have shown that Moringa oleifera is used in folk medicine to treat a variety of disorders. However, the study of immunomodulatory effects of the extracts of Moringa oleifera leaves, seeds or pods focus on anti-tumor and anti-inflammatory impacts to cell lines. Therefore, the purpose of this study was to investigate the effects of Moringa oleifera leaf extracts on systemic and mucosal Th1/Th2 cytokines secretion in murine primary cell culture. 8 to 10-week-old female BALB/c mice were used to prepare the primary cell cultures of spleen, Peyer’s patches (PP) and mesenteric lymph nodes (MLN). The spleen, PP and MLN cells were cocultured with 0, 10, 50, 100, 200, 500, and 1000μg/mL water, ethanol and ethyl acetate extracts of freezing dried and hot-wind drying Moringa oleifera leaves, lipopolysaccharide (LPS, 10μg/mL) and concanavalin A (Con A, 10μg/mL) for 24 and 48 hours. The supernatant was collected for determining IL-2, IL-4, IL-6, IL-10, IL-12p70 and IFN-gamma by ELISA. MTT assay was performed to assess the cytotoxicity of the Moringa oleifera leaf extracts. The yields of water, ethanol and ethyl acetate extracts of freezing dried Moringa oleifera leaves were 22.16±0.59 %、23.07±1.09 % and 10.47±0.46 %, respectively. The yields of boiling water, ethanol and ethyl acetate extracts of hot-wind drying Moringa oleifera leaves were 38.1± 3.96%、24.24 ± 0.08 % and 7.58 ± 2.09 %, respectively. In general, the stimulatory effects of water extracts of freezing dried Moringa oleifera leaves on the secretion of cytokine in spleen, MLN and PP cells was higher than the hot-wind drying sample. Ethyl acetate extracts did not affect the cytokines secretion of spleen, MLN and PP cells. Among the assay cytokines, the Moringa oleifera leaf extracts had the prominent effects on stimulated IL-10 secretion. Therefore, Moringa oleifera leaves may be able to promote humoral immune response through enhanced immune cells to secret IL-10.Keywords: spleen, mesenteric lymph nodes, Peyer’s patches, Moringa oleifera, cytokine, mice
Relation:	電子全文公開日期：20160910，學年度：101，70頁
Appears in Collections:	[Dept. of Health and Nutrition (including master's program)] Dissertations and Theses

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