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標題: | 黑豆免疫活性物質對人類單核細胞之免疫調節作用及其對人類白血病細胞株(U937)之生長抑制 Immunomodulation of the Active Compounds from Black Soybean [Glycine max (L.) Merr.] on Human Mononuclear Cells and the Antiproliferative Effects on U937 Cells |
作者: | Hao-Hsuan Kung 龔浩瑄 |
指導教授: | 張鴻民,吳瑞碧 |
關鍵字: | 黑豆免疫活性物質,免疫調節作用,U937細胞,生長抑制, Active Compounds from Black Soybean,Immunomodulation,U937 Cells,Antiproliferative Effects, |
出版年 : | 2006 |
學位: | 碩士 |
摘要: | 黑豆在傳統中醫藥學被認為是日常保健、醫食俱優的聖品。本實驗試著以Sephadex G-15分子篩膠體區分黑豆酒精沉澱物,並探討各區分物對人類白血病細胞U937的生長抑制情形以及對人類單核球細胞的免疫調節作用。實驗結果顯示黑豆酒精沉澱物在經過Sephadex G15分子篩膠體分離後,可得到分子量大於1500 dalton,且以醣類為主的黑豆區分一樣品(G15 P1),以及分子量1500 dalton以下之黑豆區分二樣品(G15 P2)。在細胞間接模式下,G15 P2刺激人類單核細胞後所得之條件培養液(MNC-CM)能夠有效抑制U937細胞的生長;而G15 P1則是於直接模式下對U937有明顯抑制作用。為了解G15 P1是否會誘導U937細胞凋亡亦或是透過細胞週期停滯而達到抑制功效,進一步進行U937細胞之DNA片段化實驗,以及檢測G15 P1對U937細胞週期的影響。結果發現U937並無DNA片段化的情形,但停滯於G0/G1期的比率明顯增加,故初步推測G15 P1能透過使U937細胞停滯於G0/G1期來達到抑制生長的效果。由於G15 P2能夠有效刺激人類單核細胞,並進而抑制U937細胞的生長,因此實驗第二部份便探討黑豆各階段區分物對於人類單核細胞的免疫調節作用。實驗結果顯示G15 P2能夠有效促進單核細胞的增生,增生率可達23.6 %,G15 P1則呈現些微的抑制情形;而將G15 P2刺激單核細胞後,分析其條件培養液中細胞激素的含量,亦發現G15 P2能夠有效促進人類單核細胞分泌TNF-α, IL-1β以及IFN-γ之能力。進一步利用G15 P2與人類週邊血液單核細胞培養一天後所得之條件培養液,刺激U937五天後以流式細胞儀檢測其表面抗原CD11b與CD14,其陽性表現率並無顯著增加,故推測在間接模式下G15 P2並無法誘導U937分化為單核球及巨噬細胞。 Black soybean [Glycine max (L.) Merr.] has been used as medicinal herb and food for hundreds of years. It also has been reported that black soybean contained numerous bioactive effect like radical-scavenging activity, anti-tumor activity, and activity for improving the fluidity of the whole blood. The water soluble portion of black soybean was precipitated by 75% alcohol and separated by Sephadex G-15 gel to obtain fraction 1(G15 P1) and fraction 2(G15 P2). These two fractions were used separately to stimulate human blood mononuclear cells (MNC) for one day to prepare conditioned medium (MNC-CM-1). U937 cells were incubated with condition medium for 5 days and the growth inhibition were studied. Results showed that G15 P2 stimulates human blood mononuclear cells to inhibit the cell growth of human myelocytic lymphoma U937 cells. It was also found that the MNC proliferated about 23.6 % and the MNC-CM-1 contained high levels of TNF-α, IL-1β and IFN-γ. However, in the investigation of the expression of monocyte-associated cell surface marker CD11b and CD14 on U937, we did not found differentiation of U937 cells after G15 P2 treatment. In the directly inhibitory experiments, we found G15 P1 at a dosage of 200 |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/33165 |
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顯示於系所單位: | 食品科技研究所 |
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