摘要
目的从瓦尼桑黄Sanghuangporous vaninii中分离纯化多糖,研究其结构特征及体外抗炎活性。方法采用水提醇沉法获得桑黄粗多糖(Sanghuang Polysaccharide,SHP),并利用Cellulose DE-52和Sephadex G-100色谱柱对其进行分离纯化。采用凝胶色谱—示差—多角度激光散射(gel permeation chromatography-refractive index-multi angle laser scattering,GPC-RI-MALS)、离子色谱(ion chromatography,IC)、傅里叶变换红外光谱(fourier transform infrared spectroscopy,FT-IR)、扫描电子显微镜对多糖结构进行分析;利用ELISA试剂盒测定一氧化氮(nitric oxide,NO)及炎症细胞因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和白细胞介素-1β(interleukin-1β,IL-1β)释放水平。结果从瓦尼桑黄中分离纯化得到两组均一酸性多糖SHP-1-1(0.1 mol/L NaCl)和SHP-2-1(0.2 mol/L NaCl)。SHP-1-1的重均分子量为333.599 kDa,由岩藻糖、阿拉伯糖、鼠李糖、半乳糖、葡萄糖、木糖、甘露糖、葡萄糖醛酸按照 11.48∶0.18∶0.28∶17.86∶27.71∶0.64∶11.75∶0.94 的摩尔比组成;SHP-2-1的重均分子量为563.032 kDa,由岩藻糖、阿拉伯糖、鼠李糖、半乳糖、葡萄糖、木糖、甘露糖、葡萄糖醛酸按照 0.73∶0.14∶0.32∶1.13︰14.96∶1.04∶3.79∶1.50 的摩尔比组成。SHP-1-1和SHP-2-1能够降低脂多糖诱导的RAW264.7细胞中NO、TNF-α、IL-6和IL-1β水平,其半抑制浓度为32.35~96.31μg/mL。结论SHP-1-1和SHP-2-1是具有抗炎活性的均一多糖,其结构和活性研究为瓦尼桑黄的开发和利用提供了一定参考。
Objective To isolate and purify polysaccharides from Sanghuangporus vaninii,and to investigate their structural features and in vitro anti-inflammatory activity.Methods The water-extraction and alcohol-precipitation method was used to obtain Sanghuang polysaccharide(SHP),and the Cellulose DE-52 and Sephadex G-100 columns were used for isolation and purification.The structure of the polysaccharides were analyzed using gel permeation chromatography-refractive index-multi angle laser scattering,ion chromatography,Fourier transform infrared spectroscopy,and scanning electron microscopy,and ELISA was used to observe their influence on the release of nitric oxide(NO),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β).Results Two homogeneous acidic polysaccharides,SHP-1-1 and SHP-2-1,were isolated and purified from Sanghuangporus vaninii.SHP-1-1 had a weight-average molecular weight of 333.599 kDa and was composed of fucose,arabinose,rhamnose,galactose,glucose,xylose,mannose,and glucuronic acid at a molar ratio of 11.48∶0.18∶0.28∶17.86∶27.71∶0.64∶11.75∶0.94;SHP-2-1 had a weight-average molecular weight of 563.032 kDa and was composed of fucose,arabinose,rhamnose,galactose,glucose,xylose,mannose,and glucuronic acid at a molar ratio of 0.73∶0.14∶0.32∶1.13∶14.96∶1.04∶3.79∶1.50 .Both SHP-1-1 and SHP-2-1 could reduce the levels of the inflammatory factors NO,TNF-α,IL-6,and IL-1βinduced by lipopolysaccharide in RAW264.7 cells,with a half-maximal inhibitory concentration of 32.35-96.31μg/mL.Conclusion SHP-1-1 and SHP-2-1 are homogeneous polysaccharides with anti-inflammatory activity,and the research on their structure and activity provides a certain reference for the development and utilization of Sanghuangporus vaninii.
作者
刘利娜
王仁中
吴丝雨
金传山
LIU Lina;WANG Renzhong;WU Siyu;JIN Chuanshan(Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Anhui Provincial Key Laboratory of New Manufacturing Technology for Traditional Chinese Medicine Decoction Pieces,Anhui Hefei 230012,China)
出处
《安徽中医药大学学报》
CAS
2024年第1期107-112,共6页
Journal of Anhui University of Chinese Medicine
基金
安徽中医药大学国家中药炮制传承基地项目。
关键词
瓦尼桑黄
多糖
结构表征
抗炎活性
Sanghuangporus vaninii
Polysaccharide
Structural characterization
Anti-inflammatory activity
