摘要
本文采用水提醇沉法提取美洲大蠊糖肽粗品(Periplaneta Americana glycopeptide,PAG),通过HP-20大孔树脂脱色处理PAG得脱色美洲大蠊糖肽(PAGW),DEAE-琼脂糖凝胶FF分离纯化PAGW继续得到六个组分。采用MTT法测定各个组分对巨噬细胞RAW264.7增殖能力的影响;苯酚-硫酸法及福林酚法测定PAGW中多糖和多肽的含量;PMP及OPA柱前衍生化法测定PAGW单糖和氨基酸的组成。结果显示:给药浓度范围内,PAG及PAGW随着浓度增加,均可显著升高RAW264.7的增殖活力,PAGW对其增殖率影响较好,经DEAE-琼脂糖凝胶FF洗脱的6个组分整体增殖率低。PAGW的多糖含量为21.24%,蛋白含量为76.35%,多糖由甘露糖、盐酸氨基葡萄糖等8种组成,其多肽由天冬氨酸、谷氨酸、丝氨酸、等10种氨基酸组成。结果可为美洲大蠊糖肽免疫调节活性的深入研究提供参考。
To investigate the immune regulatory activity of Periplaneta Americana glycopeptide(PAG).In this paper,Periplaneta Americana glycopeptide(PAG)was extracted by water extraction and alcohol precipitation.PAG was decolorized by HP-20 macroporous resin to obtain glycopeptide(PAGW),which was purified by DEAE-agarose gel FF to obtain six components.MTT assay was used to determine the effect of each component on RAW264.7 proliferation of macrophages.The contents of polysaccharide and protein in PAGW were determined by phenol-sulfuric acid method and folin-phenol method.The monosaccharides and amino acids of PAGW were determined by PMP and OPA pre-column derivations,respectively.The results showed that in the range of drug concentration,both PAG and PAGW could significantly increase the growth activity of RAW264.7.PAGW had a better effect on the proliferation rate of RAW264.7.DEAE-agarose gel FF eluted six components of the overall proliferation rate was low.The contents of polysaccharide and protein of PAGW are 21.24%and 76.35%,respectively.Polysaccharides are composed of 8 kinds,such as mannose and glucosamine hydrochloride.Its polypeptide are composed of 8 kinds,such as aspartic acid,glutamic acid and serine.The results can provide reference for further study on the immunomodulatory activity of Periplaneta Americana glycopeptide.
作者
王宝宇
顾婷
陈怡
袁丽萍
李开伶
肖培云
杨永寿
WANG Bao-yu;GU Ting;CHEN Yi;YUAN Li-ping;LI Kai-ling;XIAO Pei-yun;YANG Yong-shou(College of Pharmacy,Dali University,Dali 671000,China;Yunnan Provincial Key Laboratory of Insect Biopharmaceutical Research and Development,Dali 671000,China)
出处
《化学研究与应用》
CAS
CSCD
北大核心
2022年第7期1509-1516,共8页
Chemical Research and Application
基金
国家自然科学基金项目(81560634,82060747)资助。
关键词
美洲大蠊
糖肽
分离纯化
免疫活性
Periplaneta Americana
Glycopeptide
isolation and purification
immunomodulatory activity
