南柴胡多糖理化性质及拮抗白细胞黏附研究

Physicochemical properties and antagonistic capacity against cell adhesion of polysaccharide isolated from Bupleurum scorzonerifolium

目的探究南柴胡多糖(Bupleurum scorzonerifolium polysaccharide,BP)的理化性质、溶液高级构象及拮抗P-选择素介导的细胞黏附能力。方法采用水提醇沉法制备BP,利用化学法(苯酚硫酸法、间羟基联苯法、Bradford法)、高效液相色谱、傅里叶红外光谱、刚果红染色和圆二色谱分析BP的理化性质及溶液高级构象,利用细胞黏附和蛋白互作实验评价BP拮抗P-选择素介导的细胞黏附活性。结果BP是相对分子质量为36.44kDa的酸性杂多糖,糖醛酸含量为19.33%,由鼠李糖、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖5种单糖组成;BP在溶液状态下,不存在三股螺旋结构,分子具有明显的不对称性,但不对称性受浓度和温度影响;BP显著抑制P-选择素介导的HL-60与CHO-P细胞的黏附,并阻断P-选择素与P-选择素糖蛋白配体1(P-selectin glycoprotein ligand 1,PSGL-1)的相互作用。结论BP是由5种单糖组成的酸性多糖,能阻断P-选择素介导的白细胞黏附作用,具有开发成为新型抗炎药物的潜力。

Objectives To prepare Bupleurum scorzonerifolium polysaccharide(BP)to characterize its physical and chemical properties,advanced structure in solution,as well as antagonistic capacity against P-selectin-mediated cell adhesion.Methods BP was prepared by hot water extraction method,chemical methods(phenol sulfuric acid method,meta-hydroxybiphenyl method,Bradford method),high performance liquid chromatography,Fourier infrared transform spectroscopy,Congo red staining and circular dichroism were used to analyze the physical and chemical properties of BP and the advanced structure in the solution,we also evaluated its antagonistic function against P-selectin-mediated neutrophil adhesion.Results BP is an acidic heteropolysaccharide with a molecular weight of 36.44kDa.It contains 19.33%uronic acid content and is composed of five monosaccharides:Xylose,rhamnose,glucose,galactose,and arabinose.In its aqueous state,BP lacks a triple-helix structure,exhibiting pronounced asymmetry at the molecular level.However,this asymmetry is influenced by both concentration and temperature;BP significantly inhibited the P-selectin-mediated adhesion of HL-60 and Cho-P cells,and blocked the interaction between P-selectin and its physiological ligand P-selectin glycoprotein ligand 1(PSGL-1).Conclusion Bupleurum scorzonerifolium polysaccharide BP is an acidic polysaccharide composed of 5 kinds of monosaccharides,and BP can block P-selectin dependent cell adhesion,these results provide a novel therapeutical strategy for amelioration of inflammation-related disease.