枳壳均一多糖CALB-2结构表征及生物活性  

Structures Characteristics and Bioactivity of Polysaccharide CALB-2 from Aurantii Fructus

作　　者：王武静 杨燕妮 丁子禾 钟仁兴 夏天乙 李书渊 王毅 李盛青[1] 舒尊鹏 WANG Wu-jing;YANG Yan-ni;DING Zi-he;ZHONG Ren-xing;XIA Tian-yi;LI Shu-yuan;WANG Yi;LI Sheng-qing;SHU Zun-peng(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Guangdong Standardized Processing Engineering Technology Research Center of Traditional Chinese Medicine(TCM),School of TCM,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广州中医药大学中药学院,广州510006 [2]广东药科大学中药学院广东省中药饮片规范化炮制工程技术研究中心,广州510006

出　　处：《中国实验方剂学杂志》2020年第16期154-160,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(81603366);中国博士后基金面上项目(2017M610816)。

摘　　要：目的:从枳壳中分离纯化枳壳均一多糖CALB-2,并对其进行结构分析、形态表征及生物活性的研究。方法:采用热水提取法从中药枳壳中提取枳壳粗多糖(CALB),经阴阳离子交换树脂、离子交换琼脂糖凝胶以及丙烯葡聚糖凝胶等分离纯化技术得到枳壳均一多糖CALB-2;利用高效液相色谱法(HPLC)测定CALB-2的相对分子质量,采用甲基化分析和Smith降解对CALB-2进行单糖组成分析,通过红外光谱扫描(IR)分析和电镜扫描技术(SEM)对CALB-2进行结构分析及形态表征,并通过H2O2诱导心肌细胞氧化损伤模型对CALB-2抗氧化活性进行研究。结果:经HPLC测定CALB-2为均一多糖,其相对分子质量为3. 57×10^7Da。综合运用IR光谱分析、甲基化分析和Smith降解得到CALB-2为多支结构的酸性多糖,且主要以1→3,4键型存在。分子形态考察结果显示CALB-2为非晶态固体。体外活性结果表明CALB-2可明显减缓H2O2所致心肌细胞的氧化损伤作用。结论:枳壳多糖CALB-2为多支结构的均一酸性多糖,且具有一定抗心肌细胞氧化损伤的作用,以上发现为枳壳多糖进一步地深入研究奠定了理论基础。Objective:To isolate and purify a polysaccharide CALB-2 fraction from Aurantii Fructus,and analyze its basic chemical structure,morphological characteristics and bioactivity. Method: A refined CALB-2 was obtained from Aurantii Fructus by hot water extraction,then separated and purified by ion exchange resin,ion exchange agarose gel and propylene dextran gel to obtain homogeneous polysaccharide CALB-2. The molecular mass of CALB-2 was determined by high performance liquid chromatography(HPLC).Monosaccharide composition analysis of CALB-2 was conducted by methylation analysis and Smith degradation. Structural analysis and morphological characterization were conducted by infrared scanning(IR)and scanning electron microscopy(SEM)analysis. Antioxidant activity of CALB-2 was studied by using H2 O2-induced cardiomyocyte oxidative damage model. Result: CALB-2 was a homogeneous polysaccharide and the molecular weight of CALB-2 was estimated to be 3. 57×107 Da,which was proved to be a kind of highly branched acidic polysaccharides in IR analysis,methylation analysis and Smith degradation,mainly present in form of 1→3,4 bonds. Through SEM observations,we indicated that the molecular morphology of CALB-2 was amorphous solid. The in vitro activity test showed that CALB-2 had obvious protective effects on injury of H9 c2 myocardial cells induced by H2 O2. Conclusion: CALB-2 is a kind of homogeneous polysaccharide extracted from Aurantii Fructus,with an anti-cardiomyocyte oxidative damage effect,laying a theoretical foundation for further study of Aurantii Fructus polysaccharides.

关 键 词：中药枳壳 多糖CALB-2 分离纯化 结构表征 抗心肌损伤 

分 类 号：R284.2[医药卫生—中药学] R289[医药卫生—中医学]