云芝多糖B对巨噬细胞膜硫酸软骨素-蛋白聚糖结合氧化修饰低密度脂蛋白的影响

Effects of coriolus versicolor polysaccharides B on macrophage surface chondroitin sulfate-proteoglycans binding oxidized low density lipoprotein

目的探讨野生云芝多糖水溶性新组分CVPS-B对RAW264.7巨噬细胞膜硫酸软骨素-蛋白聚糖(CS-PGs)结合氧化修饰低密度脂蛋白(Ox-LDL)的影响。方法分别用溴化氰活化的Sepharose CL-4B亲和层析及体外结合CS-A的方法分别测定CS/PGs与Ox-LDL的结合率;用离子交换层析提取RAW264.7巨噬细胞膜PGs;用1,9-二甲基亚甲基蓝(DMMB)分光光度法测定糖胺聚糖。结果在17μmol MDA·g-1protein Ox-LDL水平,不同剂量CVPS-B(10、50及100μg)在体外降低Ox-LDL(200μg)与CS-A(100μg)结合,分别降低14%、29%(P<0.05)及43%(P<0.01)。不同剂量CVPS-B(10、50及100μg)在体外降低Ox-LDL(17μmol MDA·g-1 protein)与巨噬细胞膜表面PGs结合,分别降低8%、27%(P<0.05)及38%(P<0.01)。不同剂量CVPS-B可抑制Ox-LDL(50mg·L-1,17μmol MDA·g-1 protein)诱导泡沫细胞的形成。结论CVPS-B可体外抑制巨噬细胞膜CS-PGs结合Ox-LDL。

Aim To identify the effects of CVPS-B on RAW264.7 macrophage plasma membrane chondroitin sulfate （ CS ） -proteoglycans （ PGs ） binging oxidized low-density lipoprotein （Ox-LDL）. Methods Rates of PGs binding Ox-LDL were analyzed by their abilities to bind Ox-LDL coupled to a CnBr-activated Sepharose CL-4B chromatography, and rates of CS-A binding Ox- LDL were determined in vitro membrane PGs were isolated RAW264. 7 macrophages by anion exchange chromatography. Glycosaminoglycans were analyzed through the DMMB spectrophoto-metric assay. Results At the level of LDL oxidation （17 μmol MDA·g^-1 Pro）, however, CVPS-B （10, 50, 100 μg） in vitro decreased the ability of CS-A （ 100 μg ） binding Ox-LDL （200μg） by 14%, 29% （P〈0.05）, and43% （P〈0.01 ）, respectively. Furthermore, the binding of the PGs to Ox-LDL （ 17 μmol MDA·g-1 Pro） was decreased by 8%, 27% （P〈0.05）, and 38% （P〈0.01） , respectively after CVPS-B （ 10, 50, and 100 μg） being pre-incubated. CVPS-B could inhibit Ox- LDL （50 mg·L^-1 ,17 μmol MDA·g^-1 Pro）-indueed foam cell formation. Conclusions These data provide the first evidence that CVPS-B can inhibit the macrophage surface CS-PGs binding Ox-LDL in vitro.