枸杞糖缀合物和糖链对小鼠巨噬细胞功能的影响

Effects of glycoconjugate and its glycan isolated from Lycium Barbarum L on macrophage function

目的确定巨噬细胞是枸杞糖缀合物和糖链免疫作用的靶细胞之一.方法应用中性红吞噬实验和鸡红细胞吞噬实验测定了枸杞糖缀合物LbGp4和糖链LbGp4-OL对小鼠腹腔巨噬细胞吞噬功能的影响;用硝酸根还原法、酶联免疫吸附实验（ELISA）和生物活性测定法测定了巨噬细胞产生一氧化氮（NO）、IL-1β和TNF-α含量和生物活性的变化.结果 LbGp4和LbGp4-OL在（10～100）mg·L^-1剂量范围内均可剂量依赖性地促进静息巨噬细胞吞噬中性红的能力,增加活化的巨噬细胞吞噬鸡红细胞的吞噬率和吞噬指数;增加巨噬细胞培养上清NO、IL-1β和TNF-α的浓度,并增强对L929细胞的杀伤活性,促进胸腺细胞的增殖反应.结论 LbGp4和LbGp4-OL对静息和活化的腹腔巨噬细胞的吞噬功能具有明显的促进作用,对巨噬细胞产生NO、分泌IL-1β和TNF-α的含量和生物活性亦具有明显的促进作用,表明巨噬细胞是LbGp4和LbGp4-OL免疫作用的靶细胞之一.

Aim To confirm that macrophage （Mφ） is one of the immune effective target cells of glycoconjugate and its glycan isolated from the fruit of Lycium Barbarum L. Methods The phagocytic ability of peri-toneal macrophage （Mφ） was evaluated with neutral red dye phagocytosis assay and chicken red blood cells （CRBC） phagocytosis assay. Nitric oxide （NO） released by Mφ was determined with reduction reaction of NO3^-. The levels and activity of interleukin-1β （ IL- 1β） and tumor necrosis factor-α（TNF-α） secreted by Mφ were measured using enzyme linked immunoassay （ELISA） and biological activity assay. Results LbGp4 and LbGp4-OL（ 10 - 100） mg · L^-1 increased the contents of neutral red dye phagocytized by resting Mφ and the CRBC phagocytosis index of starch activated Mφ was also elevated. The levels of NO, IL-1β and TNF-α produced by resting Mφ were promoted after incubation of resting Mφ with LbGp4 or LbGp4-OL. The biological activity of IL-1β and TNF-α were augmented toward L929 cell and mouse thymocyte as target cells respectively. Conclusions LbGp4 and LbGp4OL enhanced the phagocytic and secretocytic functions of Mφ,which suggested that Mφ is also the main immune effective target cell of LbGp4 and LbGp4-OL.