权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Polysaccharides form mushroom stimulate macrophages through intestinal epithelial cells

蘑菇中的多糖通过肠上皮细胞刺激巨噬细胞

基本信息

批准号：
16500509
负责人：
MIZUNO Masashi
金额：
$ 2.37万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

Lentinan has been known as an antitumor polysaccharide contained in Lentinula edodes, and its molecular weight was estimated to be more than 400,000. Lentinan shows the antitumor activity through host-mediated, because it does not kill cancer cells. However, it has not been ascertained whether or not lentinan was absorbed in intestine. In this study, co-culture system using intestinal epithelial cell line, caco2 cells (apical side), and macrophage cell lines, RAW 264.7 cells (basolateral side), was applied to clarify how lentinan stimulated macrophage. Caco2 cells (2.5×10^5 cells/well) were treated with 50 μg/ml lentinan, and then were co-cultured with RAW 264.7 cells (5×10^5 cells/well). TNF-α production was detected just in basolateral side (150 pg), not in apical side. When catalase was added in basolateral side in the same condition, TNF-α production was inhibited to 50%, but superoxide dismutase did not. RAW 264.7 cells stimulated directly with lentinan did not produce TNF-α. Furthermore, when caco2 cells were cultured with lentinan at the same concentration without RAW 264.7 cells, they did not produce TNF-α in both sides. Neither lentinan was detected in basolateral side. These findings suggested that intestinal epithelial cells stimulated with lentinan produced H_2O_2 and newly produced H_2O_2 played an important role in cell to cell cross talking to enhance TNF-α production from macrophages.

香菇多糖被认为是香菇中含有的一种抗肿瘤多糖，其分子量估计超过40万。香菇多糖通过宿主介导表现出抗肿瘤活性，因为它不会杀死癌细胞。然而，尚未确定香菇多糖是否被肠道吸收。在本研究中，应用使用肠上皮细胞系caco2细胞（顶端侧）和巨噬细胞细胞系RAW 264.7细胞（基底外侧）的共培养系统来阐明香菇多糖如何刺激巨噬细胞。 Caco2细胞（2.5×10^5细胞/孔）用50 μg/ml香菇多糖处理，然后与RAW 264.7细胞（5×10^5细胞/孔）共培养。仅在基底外侧（150 pg）而非顶端检测到 TNF-α 的产生。当在相同条件下在基底外侧添加过氧化氢酶时，TNF-α的产生被抑制至50%，但超氧化物歧化酶则没有。直接用香菇多糖刺激的 RAW 264.7 细胞不产生 TNF-α。此外，当caco2细胞与相同浓度的香菇多糖一起培养而没有RAW 264.7细胞时，两侧均不产生TNF-α。基底外侧均未检测到香菇多糖。这些发现表明，用香菇多糖刺激的肠上皮细胞产生的H_2O_2和新产生的H_2O_2在细胞间的交叉对话中发挥重要作用，从而增强巨噬细胞产生TNF-α。