水龙骨多糖成分及免疫调节活性的初步研究

目的研究水龙骨多糖(Rhizoma polypodiodis nipponicae polysaccharide,RPNP)的组成及体内外免疫调节活性。方法采用水提醇沉法提取RPNP,HPLC分析单糖组成,MTT法考察RPNP对Raw264.7细胞增殖能力的影响,中性红染色法考察其对Raw264.7细胞吞噬能力的影响,ELISA检测细胞因子的分泌水平。采用环磷酰胺诱导建立免疫抑制小鼠模型,考察RPNP对其脏器指数和小肠派氏结数量的影响。结果RPNP是由Man、Rha、Glc、Gal、Ara5种单糖组成的酸性杂化多糖,其总糖、糖醛酸、蛋白质及核酸含量分别为42.98%、25.25%、1.30%、5.87%。适当剂量的RPNP能够提高Raw264.7细胞的增殖及吞噬能力,促进TNF-α及IL-6的分泌,并能够增加免疫抑制小鼠的脏器指数及派氏结个数。结论从水龙骨中分离得到的酸性杂化多糖,在适当剂量下能够活化巨噬细胞,增强免疫抑制小鼠免疫功能。

The composition and immunomodulatory activity of polysaccharide from Rhizoma polypodiodis nipponicae

This study was performed to investigate the composition and immunomodulatory activity of Rhizoma polypodiodis nipponicae polysaccharide (RPNP).The RPNP was extracted by hot water and precipitated in ethanol, and then monosaccharide composition was analyzed by HPLC.MTT was used to evaluate the effects of RPNP on the proliferation of Raw264.7 cell;neutral red staining method was employed to investigate the effects of RPNP on the phagocytosis of Raw264.7 cell,while ELISA was used to detect the effects of RPNP on the secretion of cytokines. Furthermore,the immunomodulatory activity of RPNP in vivo was evaluated using immunosuppressive mice model induced by cyclophosphamide (CY).Data showed that RPNP was an acidic polysaccharide composed of Man,Rha, Glc,Gal and Ara.The contents of total carbohydrate,glucuronic acid,protein and nucleic acid were 42.98%, 25.25%,1.30% and 5.87%,respectively.RPNP could increase the proliferation and phagocytosis of Raw264.7 cells dose-dependently,and promoted the secretion of TNF-α and IL-6.RPNP also increased the organ index and the number of Peyer's patches in immunosuppressive mice.In conclusion,RPNP can enhance the immune function of macrophages and protect mice against immunosuppression induced by CY.