白芍多糖提取工艺优化、分子特性分析及其对巨噬细胞的增殖作用

Molecular Characteristics of Polysaccharides from Radix Paeoniae Alba and Its Effects on Macrophages

本文采用水提醇沉法提取白芍多糖,通过正交试验确定白芍多糖提取工艺参数并分析其化学组成,使用高效尺寸排阻色谱-紫外检测器-多角度激光光散射仪-示差折光检测器联机系统检测多糖分子量,利用气相色谱-质谱联用仪分析多糖的单糖组成和链接方式,采用WST法检测白芍多糖对巨噬细胞RAW264.7增殖率的影响。结果表明:白芍多糖的最佳提取工艺参数为液料比25:1 mL/g、提取温度85℃、提取时间3.0 h,得率12.40%。白芍多糖含有96.2%±1.2%总糖和5.5%±0.6%蛋白质,未检出硫酸根和糖醛酸,其分子质量和回转半径分别为2.3×10^(6) u和234.9 nm。白芍多糖主要含葡萄糖,大部分是通过(1→4)糖苷键链接的。同时,白芍多糖可以在2、5、10μg/mL浓度范围促进RAW264.7细胞增殖。其中,10μg/mL处理组的细胞增殖率得到大幅提升,且与其他组别的细胞增殖率存在显著性差异(P<0.05)。

The polysaccharide was extracted from Radix Paeoniae Alba by the method of water extraction and ethanol precipitation. The optimum extraction parameters of the polysaccharide were determined by orthogonal test and its chemical compositions was investigated. The molecular weight of the polysaccharide was analyzed by HPSEC-UV-MALLS-RI system, which monosaccharide composition and glycosidic linkage was investigated by GC-MS. The effect of the polysaccharides on RAW264.7 cells proliferation was determined by WST assay. The results showed that the optimum extraction parameters were as follows: the ratio of liquid to material was 25:1 mL/g, extraction temperature 85 ℃,extraction time 3.0 h, with the yield of 12.40%. The polysaccharide was composed of 96.2%±1.2% total sugar and5.5%±0.6% protein, no sulfate and uronic acid, with the molecular weight and radius of gyration 2.3×10^(6) u and 234.9 nm,respectively. The polysaccharide mainly contained glucose with the linkage of →4)-Glu-(1→, which also could promote the proliferation of RAW264.7 cells at the concentrations of 2, 5, 10 μg/mL.It was nothing that the cell proliferation rate of the10 μg/mL treatment group was significantly increased, which exhibitedsignificant difference with other groups(P< 0.05).