【摘要】 目的:从猪脑中提取分离糖胺聚糖（glycosaminoglycans,GAGs）,并进行精细结构分析。方法:猪脑组织经脱脂、蛋白酶酶解后,经氯代十六烷基吡啶（CPC）富集,乙醇沉淀获得糖胺聚糖粗品。利用Q-Sepharose Fast Flow离子交换柱纯化,获得7个组分（P1～P7）。所得分级样品分别经肝素裂解酶与硫酸软骨素裂解酶酶解,2-氨基吖啶酮（AMAC）荧光标记后,利用液质联用方法分析其精细结构。结果:猪脑GAGs粗品中主要获得硫酸乙酰肝素（heparan sulfate,HS）、硫酸软骨素（chondroitin sulfate,CS）和少量透明质酸（hyaluronic acid,HA）,含量分别为307,293和18 mg·kg^-1。P2～P4组分GAGs含量较少; P5～P7组分CS和HS含量相当,CS中以ΔUA-GalNAc4S为主; HS组成以ΔUA-GlcNAc为主,电荷密度略低,未获得肝素组分。结论:基于液质联用技术分析猪脑组织的精细结构,为动物来源糖胺聚糖的研究提供依据。更多还原

【Abstract】 Objective: To extract and purify glycosaminoglycans （GAGs） from porcine brain and analyze their fine structures. Methods: Homogenized porcine brain was defatted then digested with trypsin and papain. The crude GAGs were enriched by cetylpyridinium chloride （CPC） and precipitated by ethanol. Seven fractions were eluted by Q-Sepharose Fast Flow column and named as P1 ～ P7,respectively. The crude GAGs and purified fractions were digested by heparin lyase I/II/III and chondroitin lyase ABC,respectively. The generated disaccharides were labeled with 2-AMAC and then analyzed by LC-MS. Results: The GAGs isolated from porcine brain were mainly composed of heparan sulfate （HS） and chondroitin sulfate （CS） with trace amount of hyaluronic acid （HA）,and their content were 307,293 and 18 mg·kg^-1,respectively. GAGs were less in P2 ～ P4,and P5 ～ P7 had similar contents of CS and HS. CS from P5 ～ P7 were all primarily composed of ΔUA-GalNAc4S. And ΔUAGlcNAc was the main disaccharide detected in HS,which had a lower charge density. No heparin was acquiredfrom porcine brain. Conclusions: This study evaluated the contents and exact composition of GAGs in porcine brain based on LC-MS. Detailed characterization of the disaccharide structures and compositions is beneficial for study on animal-derived GAGs.更多还原