石耳与黑石耳多糖的抗炎作用研究

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...

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

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^-1lipopolysaccharide. After adding 5 μg·mL^-1HUEP 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^-1ZDMP 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 for polysaccharides from Umbilicaria esculenta or Dermatocarpon miniatum, and then polysaccharides from Dermatocarpon miniatum exhibit better anti-inflammation effects than another one.