黑螵蛸多糖的制备及活性测定
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摘要
目的对黑螵蛸多糖进行分离纯化,并初步测定其活性。方法采用水提-醇沉法制备黑螵蛸粗多糖,经DEAE Sephadex A-25离子交换柱色谱和Sephadex G-75凝胶柱色谱纯化得到均一的黑螵蛸多糖组分HPP-1。采用凝胶渗透色谱,紫外光谱等技术对HPP-1进行分析,另外探究多糖的抗氧化活性、抗菌活性和胆碱酯酶抑制活性。结果黑螵蛸多糖HPP-1的相对分子质量(Mr)为13 048,不含蛋白质、核酸和色素,DPPH自由基和羟基自由基清除IC_(50)值分别为10.35±0.12,100.33±0.06μg/ml;多糖对枯草芽孢杆菌有抑制活性,最小抑菌浓度为18.8μg/ml;抑制乙酰胆碱酯酶和丁酰胆碱酯酶的IC_(50)值分别为6.52±0.03,18.26±0.26μg/ml。结论黑螵蛸多糖具有较好的抗氧化活性、抗菌活性和胆碱酯酶抑制活性,可作为潜在的抗氧化剂、抑菌剂和胆碱酯酶抑制剂进行深入研究和开发。
Objective To isolate, purifficate and preliminarily determine the activity of the polysaccharide from Mantidis O?theca(Heipiaoxiao). Methods The crude polysaccharide was obtained by hot water extration and ethanol deposition, and the polysaccharide(HPP-1) was purified using DEAE Sephadex A-25 and Sephadex G-75 column chromatography. The HPP-1 was elucidated by GPC and UV. In addition, the antioxidant activity, antibacterial activity and cholinesterase inhibitory activity of the polysaccharide were explored. Results The relative molecular weight(Mr)of the polysaccharide HPP-1 from Mantidis O?theca(Heipiaoxiao) was 13 048. It was free of protein, nucleic acid and pigments. IC_(50) values of DPPH and hydroxyl radicals scavenging in vitro were 10.35±0.13, 100.33±0.06 μg/ml, respectively. HPP-1 had inhibitory activity against Bacillus subtilis, with a minimum inhibitory concentration of 18.8 μg/ml. The IC_(50) values for inhibition of AChE and BuChE were 6.52±0.03, 18.26±0.26 μg/ml, respectively.Conclusion Mantidis O?theca(Heipiaoxiao) polysaccharide has good antioxidant activity, antibacterial activity and cholinesterase inhibitory activity, and may be found promising applications as potent antioxidant agent, bacteriostatic agent and double cholinesterase inhibitors.
Objective To isolate, purifficate and preliminarily determine the activity of the polysaccharide from Mantidis O?theca(Heipiaoxiao). Methods The crude polysaccharide was obtained by hot water extration and ethanol deposition, and the polysaccharide(HPP-1) was purified using DEAE Sephadex A-25 and Sephadex G-75 column chromatography. The HPP-1 was elucidated by GPC and UV. In addition, the antioxidant activity, antibacterial activity and cholinesterase inhibitory activity of the polysaccharide were explored. Results The relative molecular weight(Mr)of the polysaccharide HPP-1 from Mantidis O?theca(Heipiaoxiao) was 13 048. It was free of protein, nucleic acid and pigments. IC_(50) values of DPPH and hydroxyl radicals scavenging in vitro were 10.35±0.13, 100.33±0.06 μg/ml, respectively. HPP-1 had inhibitory activity against Bacillus subtilis, with a minimum inhibitory concentration of 18.8 μg/ml. The IC_(50) values for inhibition of AChE and BuChE were 6.52±0.03, 18.26±0.26 μg/ml, respectively.Conclusion Mantidis O?theca(Heipiaoxiao) polysaccharide has good antioxidant activity, antibacterial activity and cholinesterase inhibitory activity, and may be found promising applications as potent antioxidant agent, bacteriostatic agent and double cholinesterase inhibitors.
引文
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[3]葛德燕,陈祥盛.桑螵蛸药用历史与研究进展[J].山地农业生物学报,2006,25(5):455-460.
[4]贾坤静,艾雪,贾天柱,等.桑螵蛸炮制前后蛋白质和多糖及脂类成分比较[J].亚太传统医药,2015,11(23):15-17.
[5]魏暑飓,何江波,晏永明.桑螵蛸植物源化学成分的研究[J].药学研究,2013,32(3):128-130.
[6]林璐璐,牛长缨,雷朝亮.桑螵蛸及其粗提物对四氧嘧啶糖尿病小鼠的影响[J].时珍国医国药,2009,20(8):1901-1903.
[7]王文东,郭雪芳,刘颖丽,等.桑螵蛸挥发油对金黄色葡萄球菌生物被膜形成的体外抑制效应的初步探究[J].中国微生态学杂志,2015,27(11):1250-1252.
[8]Wei W,Feng L,Nie S P,et al.Structure characterization and immunomodulating effects of polysaccharides isolated from Dendrobium officinale[J].J Agr Food Chem,2016,64(4):881-889.
[9]Chen H,Zhang M,Qu Z,et al.Compositional analysis and preliminary toxicological evaluation of a tea polysaccharide conjugate[J].J Agr Food Chem,2007,55(6):2256-2260.
[10]Chen G,Yuan Q,Saeeduddin M,et al.Recent advances in tea polysaccharides:Extraction,purification,physicochemical characterization and bioactivities[J].Carbohyd Polym,2016,153(22):663-678.
[11]常双艳,蓝炎阳,王少峰.多糖提取纯化方法及其生物活性研究进展[J].福建热作科技,2013(4):34-38.
[12]Romdhane M B,Haddar A,Ghazala I,et al.Optimization of polysaccharides extraction from watermelon rinds:Structure,functional and biological activities[J].Food Chem,2017,216(56):355-364.
[13]宋波,李宗阳,刘亚旻,等.独活抑制乙酰胆碱酯酶和丁酰胆碱酯酶活性研究[J].中南药学,2011,9(10):721-724.
[14]曾婷,彭梅,杨娟.仙茅多糖的分离纯化及结构分析[J].中国实验方剂学杂志,2016,22(16):53-57.
[15]王琳炜,欧阳臻,张碧娟,等.霍山铁皮石斛多糖的脱蛋白工艺及结构分析[J].食品科学,2017,38(12):164-170.
[16]大连轻工业学院.食品分析[M].北京:中国轻工业出版社,1994:177-180
[17]尚晓娅,任锦,曹刚,等.山药多糖的制备及其体外抗氧化活性[J].化学研究,2010,21(2):72-76.
[18]朱璐,董福,冯叙桥,等.浸提法、超声波法和微波法提取紫薯花色苷的抗氧化性比较研究[J].食品科学,2015,36(19):83-88.
[19]郑宇翔,肖凤霞,林励,等.青蒿渣中总多糖提取工艺及其抗氧化活性[J].中国实验方剂学杂志,2015,21(14):8-11.
[20]吕毅.海蛙(Fejervarya cancrivora)皮肤分泌液中抗菌多肽的分离纯化、分子克隆以及结构与功能的研究[D].南京:南京农业大学,2009.
[21]Joubert J,Foka G B,Repsold B P,et al.Synthesis and evaluation of 7-substituted coumarin derivatives as multimodal monoamine oxidase-B and cholinesterase inhibitors for the treatment of Alzheimer’s disease[J].Eur J Med Chem,2017,125(41):853-864.
[22]曾维才,石碧.天然产物抗氧化活性的常见评价方法[J].化工进展,2013,32(6):1205-1213.