摘要
对海洋真菌(Phoma herbarum YS4108)多糖YCP进行羧甲基化修饰,研究其产物(YCPC)的理化性质及其对巨噬细胞免疫活性的影响。采用氢氧化钠—氯乙酸法对YCP进行羧甲基化修饰,通过红外和核磁光谱法对产物进行鉴定,在可见光谱范围检测其溶液透光率,Griess法测定其诱导巨噬细胞分泌NO的活性,并通过流式细胞仪检测其结合巨噬细胞的能力。结果:YCP羧甲基化修饰后得到了10种不同取代度的YCPC样品,红外光谱的COO-峰及13C核磁光谱的C=O峰归属于引入的羧甲基;YCPC的水溶液的透光率:YCPC6>YCPC10>YCP;诱导巨噬细胞分泌NO的能力:YCP>YCPC10>YCPC6;且YCPC对YCP结合小鼠腹腔巨噬细胞有竞争性抑制作用。YCP羧甲基化修饰后,产物的水溶液透光率明显提高,并具有和YCP相同的巨噬细胞表面特异性结合位点,YCPC诱导巨噬细胞分泌NO的能力随取代度升高而下降。
To prepare the carboxymethylated productions of YCP, which were named YCPC, and research on the physicochemical properties and immunological activity of YCPC. YCP was carboxymethylated by the chloroacetic acid--NaOH method. Structure confirmation of YCPC was identified by IR and NMR. The transmittance of polysaccharide water solution was determined by method of visual full wavelength scanned. Production of NO induced by macrophages was measured by method of Griess. The capability binding to mouse peritoneal macrophages was subjected to flow cytometric analysis. Experiment results showed that the ten products of carboxymethylation were obtained with different substitute degrees （DS）. The new bands showed by IR spectrometry and a peak showed by ^13C NMR spectra were attributed to the group of CH3 COO^-, which is identified to be the structure of YCPC. The transmittance of polysaccharide water solutions was obtained in the following order： YCPC6〉YCPC10〉YCP. The amount of NO stimulated by the polysaccharides followed this order： YCP〉YCPC10〉YCPC6. YCPC could compete wtih YCP-FLA for binding to mouse peritoneal macrophages. Transmittance of YCPC water solutions was improved compared to that of YCP. YCPC may share the similar specific binding sites on mouse macrophages with YCP. The amount of NO decreased when YCPC was with the higher DS.
出处
《药物生物技术》
CAS
CSCD
2009年第4期369-373,共5页
Pharmaceutical Biotechnology
基金
国家高技术研究发展计划(863计划)资助项目(NO.2006AA09302)
国家自然科学基金资助项目(NO.30672479
30873201)
高等学博士学科点专项科研基金(NO.20060316001)
关键词
海洋真菌多糖YCP
羧甲基化
透光率
免疫活性
Marine fungus polysaccharide YCP, Carboxymethylation, Transmittance, Immunological activity
