Stimulation of phagocytosis and phagosome-lysosome fusion by glycolipids from Grifola frondasa (Maitake).

灰树花 (舞茸) 的糖脂刺激吞噬作用和吞噬体-溶酶体融合。

基本信息

批准号：
08680067
负责人：
MIYAZAKI Yoshiko
金额：
$ 0.83万
依托单位：
Mukogawa Women's University Junior College Division
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

In this report, I attempted to elucidate the relationship between the structure of glycolipids from Grifola frondosa (Maitake) and the mechnism of phagocytic processes in human monocytes (MN) or polymorphonuclear leukocytes (PMN). The characteristic lipid components of Grifola frondosa (Maitake) were cardiolipin (CL), phosphatidylserine (PS) phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidic Acid (PA), phosphatidylcholine (PC), phosphatidyl-ethanolamine (PE), and three classes of glycolipids (MGL-1,2,3). The structures of glycolipids from Maitake were determined by thin layr chromatography (TLC), Infrared spectrometry, FAB mass spectrometry and NMR spectrometry. The effect of glycolipids on phagocytosis by human PMN or MN using killed Staphylococcus aureus cells coated with glycolipids (MGL-1,2,3) was examined. The rate of phagosome-lysosome fusion by PMN was determined by counting acridine orange-stained bacteria under a fluorescence microscope. Superoxide anion production was determined by the reduction of cytochrome c. Both phagocytosis and phagosome-lysosome fusion of MGL-2 was stimulated in dose-dependent manner, while MGL-1 and MGL-3 did not. Relative phagosome-lysosome fusion rate of MGL-2-coated staphylococci was six times higher than that of the noncoated control and phagocytic rate was twice, respectively. The superoxide anion release from PMN was not affected by MGL-2. I compared the effect of lipopolysaccharide (LPS) from Salmonella enteritidis with that of MGL-2 from Maitake. The glycolipid (MGL-2) reversed the inhibition of phagocytosis by LPS.

在这份报告中，我试图阐明灰树花（舞茸）糖脂结构与人单核细胞（MN）或多形核白细胞（PMN）吞噬过程机制之间的关系。灰树花 (Maitake) 的特征性脂质成分为心磷脂 (CL)、磷脂酰丝氨酸 (PS)、磷脂酰胆碱 (PC)、磷脂酰肌醇 (PI)、磷脂酸 (PA)、磷脂酰胆碱 (PC)、磷脂酰乙醇胺 (PE) 和三种糖脂类别 (MGL-1,2,3)。通过薄层色谱法（TLC）、红外光谱法、FAB 质谱法和核磁共振波谱法测定舞茸糖脂的结构。使用涂有糖脂（MGL-1,2,3）的灭活金黄色葡萄球菌细胞，检查了糖脂对人 PMN 或 MN 吞噬作用的影响。通过在荧光显微镜下计数吖啶橙染色的细菌来测定PMN的吞噬体-溶酶体融合率。超氧阴离子的产生通过细胞色素c的还原来测定。 MGL-2 的吞噬作用和吞噬体-溶酶体融合均以剂量依赖性方式受到刺激，而 MGL-1 和 MGL-3 则不然。 MGL-2包被的葡萄球菌的相对吞噬体-溶酶体融合率比未包被的对照高六倍，吞噬率分别是两倍。 PMN 中超氧阴离子的释放不受 MGL-2 的影响。我比较了肠炎沙门氏菌脂多糖 (LPS) 与舞茸 MGL-2 的作用。糖脂（MGL-2）逆转了LPS对吞噬作用的抑制。