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

在这份报告中，我试图阐明灰树花糖脂的结构与人单核细胞(MN)或多形核白细胞(PMN)吞噬过程的机制之间的关系。灰树花的特征脂成分为心磷脂(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(MGL-1、2、3)。采用薄层层析(TLC)、红外光谱、原子吸收光谱、核磁共振谱等方法对其糖脂进行了结构鉴定。用包被糖脂(MGL-1、2、3)的金黄色葡萄球菌灭活细胞，研究了糖脂对人PMN或MN吞噬作用的影响。在荧光显微镜下计数吖啶橙染色菌，测定PMN吞噬小体-溶酶体融合率。MGL-2的吞噬作用和吞噬小体-溶酶体融合均呈剂量依赖性刺激，而MGL-1和MGL-3则无此作用。MGL-2包被葡萄球菌的相对吞噬小体-溶酶体融合率是未包被对照的6倍，吞噬百分率是未包被对照的2倍。MGL-2对PMN释放超氧阴离子无明显影响。我比较了肠炎沙门氏菌的脂多糖(LPS)和Maitake的MGL-2的作用。糖脂(MGL-2)可逆转内毒素抑制的吞噬作用。

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.