College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China; School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Int J Biol Macromol. 2021 Jun 1;180:129-142. doi: 10.1016/j.ijbiomac.2021.03.006. Epub 2021 Mar 5.
The current experiments were designed to explore the structural features and immunopotentiation activity of two homogeneous polysaccharides PCSPA and PCSPB prepared from Crocus sativus petals using DEAE-Sephadex A-50 and Sephadex G200 column chromatography. The structures of PCSPA and PCSPB were systematically characterized using extensive chemical and spectroscopic methods including colorimetry, HPGPC-RID, GC-MS, Smith degradations, methylation, solvolytic desulfation, UV, FT-IR, NMR, SEM, and AFM. The average molecular weights of PCSPA and PCSPB were 1.98 × 10 and 2.53 × 10 Da, respectively. PCSPA consisted of Gal, Rha, Ara, and Xyl in the molar ratio of 16:5:7:3, while PCSPB were composed of Gal, Glc, Man, Rha, Ara, and Xyl with molar ratio of 16:2:7:19:15:16. Both polysaccharides contained sulfonic and acetyl groups. PCSPA and PCSPB significantly activated RAW264.7 cells by enhancing the phagocytic activity, up-regulating the expression of surface molecules, promoting the production and mRNA expression of cytokines and chemokines via MAPK and NF-κB pathway.
本研究旨在通过 DEAE-Sephadex A-50 和 Sephadex G200 柱层析，从番红花花瓣中提取两种均一多糖 PCSPA 和 PCSPB，并对其结构特征和免疫增强活性进行研究。采用比色法、HPGPC-RID、GC-MS、Smith 降解、甲基化、溶剂解脱硫、UV、FT-IR、NMR、SEM 和 AFM 等广泛的化学和光谱方法系统地表征了 PCSPA 和 PCSPB 的结构。PCSPA 和 PCSPB 的平均分子量分别为 1.98×10 和 2.53×10 Da。PCSPA 由 Gal、Rha、Ara 和 Xyl 组成，摩尔比为 16:5:7:3，而 PCSPB 由 Gal、Glc、Man、Rha、Ara 和 Xyl 组成，摩尔比为 16:2:7:19:15:16。两种多糖均含有磺酸基和乙酰基。PCSPA 和 PCSPB 通过增强 RAW264.7 细胞的吞噬活性，上调表面分子的表达，促进细胞因子和趋化因子的产生和 mRNA 表达，从而显著激活 RAW264.7 细胞，该过程涉及 MAPK 和 NF-κB 通路。
