石耳与黑石耳多糖的抗炎作用研究  

Polysaccharides with Anti-Inflammatory Properties from Umbilicaria esculenta and Dermatocarpon Miniatum

作　　者：王昭晶[1] 曹森[1] WANG Zhao-jing;CAO Sen(Department of Bioengineering and Biotechnology,Huaqiao University,Xiamen 361021,China)

机构地区：[1]华侨大学生物工程与技术系,福建厦门361021

出　　处：《中国药学杂志》2022年第1期24-29,共6页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(31201314)。

摘　　要：目的对4个不同产地石耳和黑石耳多糖的理化性质、微观结构和抗炎活性进行研究。方法采用热水提取法提取粗多糖。利用Sevag法,透析和DEAE-Sepharose CL-6B柱色谱等方法进行分离纯化。高效体积排阻色谱法和红外光谱法等研究理化性质,透射电子显微镜观察微观结构,实时定量PCR研究抗炎症作用。结果得到黄山石耳多糖(HUEP),宁安石耳多糖(NUEP),张家界黑石耳多糖(ZDMP)和神农架黑石耳多糖(SDMP),得率分别为质量分数20%,15.40%,48%和38.25%。HUEP和NUEP由均一多糖组成,ZDMP和SDMP由不均一糖聚合而成。4种纯化多糖均由葡萄糖、木糖、半乳糖和甘露糖组成,HUEP和NUEP各单糖占比均匀,ZDMP和SDMP葡萄糖比例高。HUEP呈现无分支的长链交织结构,NUEP由3股以上无分支的短链缠绕组成,ZDMP和SDMP则由长短不一的链聚合组成。采用1μg·mL^(-1)脂多糖诱导小鼠巨噬细胞RAW264.7建立炎症模型,添加5μg·mL^(-1)的HUEP或NUEP,结果显著降低白细胞介素1βmRNA的表达(抑制率HUEP=89.06%,NUEP=84.73%,P<0.000 1)。添加75μg·mL^(-1)的ZDMP或SDMP则能显著降低所有炎症因子的mRNA表达,且部分抗炎效果高于阳性对照地塞米松。结论产地不同对石耳或黑石耳多糖的特性和抗炎活性没有影响,黑石耳多糖抗炎效果优于石耳多糖。OBJECTIVE To study physicochemical properties, microstructures and anti-inflammation effects of polysaccharides from Umbilicaria esculenta and Dermatocarpon miniatum coming from four different fields. METHODS Crude polysaccharides were isolated using water extraction method and purified with Sevag, dialysis and DEAE-Sepharose CL-6 B. Physicochemical properties were analyzed following methods of high-performance size-exclusion chromatography and Fourier transform infrared spectroscopy, microstructures were observed with transmission electron microscopy, and anti-inflammation activities were identified using real time quantitative PCR. RESULTS Four kinds of major polysaccharides The Huangshan Umbilicaria esculenta polysaccharide(HUEP),Ningan Umbilicaria esculenta polysaccharide(NUEP),Zhangjiajie Dermatocarpon miniatum polysaccharide(ZDMP) and Shennongjia Dermatocarpon miniatum polysaccharide(SDMP) were purified with yields of 20%, 15.40%, 48% and 38.25%, respectively. HUEP and NUEP were homogeneous, while ZDMP and SDMP were not. They consisted mainly of glucose, xylose, galactose and mannose, and four monosaccharides were even distribution in HUEP and NUEP, while ZDMP and SDMP were composed of the high percentage of glucose. Through TEM, long linear molecules were observed in HUEP and short linear molecules with three entangled chains were identified in NUEP,while both long and short chains were obtained in ZDMP and SDMP molecules. Mouse macrophage cells RAW264.7 were induced to produce inflammatory cytokines(TNF-α, IL-6, IL-1β and COX-2) by adding 1 μg·mL^(-1) lipopolysaccharide. After adding 5 μg·mL^(-1) HUEP or NUEP in LPS-induced cells, IL-1β mRNA expression levels were significantly reduced(HUEP=89.06%,NUEP=84.73%, P<0.000 1), while the mRNA expression levels of four inflammatory factors were reduced with the addition of 75 μg·mL^(-1) ZDMP or SDMP and some of the effects were more than dexamethasone. CONCLUSION Different fields have no influence in the terms of properties and anti-inflammation effects

关 键 词：石耳 黑石耳 微观结构 抗炎症作用 

分 类 号：R282[医药卫生—中药学]