首页|桑叶多糖结构鉴定及其降血糖活性研究

桑叶多糖结构鉴定及其降血糖活性研究

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目的 阐明桑叶中具有降血糖活性的多糖结构基础,为桑叶的开发和利用提供理论基础。方法 采用水提醇沉法获得桑叶粗多糖,经离子交换色谱和凝胶渗透色谱分离纯化获得SY02、SY02-3、SY02-3A及SY02-3B等多糖组分,并测定均一多糖的理化性质;联合单糖组成分析、糖残基连接方式分析和核磁分析对桑叶多糖进行结构鉴定;采用棕榈酸(Palimitate,PA)诱导INS-1细胞凋亡模型、Caspase3/7活性试剂盒测定桑叶多糖对INS-1E细胞凋亡的保护作用,并运用Western blot法检测桑叶多糖对INS-1E细胞凋亡关键蛋白Cleaved Caspase 3表达的影响。结果 采用水提醇沉法获得桑叶粗多糖SY,经离子交换树脂柱分离再由凝胶柱纯化得到SY02-3,再次纯化后得到均一酸性多糖SY02-3A与蛋白多糖SY02-3B,两者分子量分别为3。7×104 Da和1。03×104 Da。单糖组成分析结果表明SY02-3A含有鼠李糖、半乳糖醛酸、半乳糖、木糖、阿拉伯糖,摩尔比分别为23。97∶33。85∶11。73∶5。04∶25。41,得率为0。32%;SY02-3B由鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、木糖与阿拉伯糖组成,其摩尔比19。83∶10。34∶45。42∶9。01∶2。23∶13。17,得率为2。00%;SY02-3B含有14种氨基酸,其中天冬氨酸、谷氨酸、丙氨酸和甘氨酸含量相对较高。生物活性研究表明SY02-3可以保护PA诱导的INS-1E细胞凋亡,这种作用可能与降低 Caspase 3 活性和下调 cleaved Caspase 3 蛋白表达有关。结论 桑叶中含有酸性肽聚糖(SY02-3A)和蛋白聚糖(SY02-3B)的桑叶多糖SY02-3具有抑制PA诱导的INS-1E细胞凋亡的作用。
Structure Identification and Hypoglycemic Activity of Polysaccharides from Mulberry Leaves
Objective To elucidate the structural basis of polysaccharides with hypoglycemic activity in mulberry leaves,and provide a theoretical basis for the development and utilization of mulberry leaves.Methods The crude polysaccharides from mulberry leaves were extracted by hot water.The components of SY02,SY02-3,SY02-3A and SY02-3B were obtained by ion-exchange chromatography and gel permeation chromatography.We determined the physicochemical properties of homogeneous polysaccharide.The structure of mulberry leaf polysaccharide was identified by monosaccharide composition analysis,sugar residue linkage analysis and nuclear magnetic analysis.Palimitate(PA)induced INS-1 cell apoptosis model was used to determine the protective effect of mulberry leaf polysaccharide on INS-1E cell apoptosis,and Caspase3/7 activity kit was used to detect mulberry leaf polysaccharide on the key protein of INS-1E cell apoptosis Cleaved by Western blot method effect of Caspase 3 expression.Results The crude polysaccharide SY was extracted from mulberry leaves by hot water,which was separated by ion exchange resin column and purified by gel column to obtain SY02-3.After purification,we obtained homogeneous polysaccharide SY02-3A and proteoglycan SY02-3B,with molecular weights of 3.7×104 Da and 1.03×104 Da.The results of monosaccharide composition analysis showed that SY02-3A contained rhamnose,galacturonic acid,galactose,xylose and arabinose.The molar ratio of SY02-3A was 23.97:33.85:11.73:5.04:25.41,and the yield was 0.32%.SY02-3B is composed of rhamnose,galacturonic acid,glucose,galactose and arabinose.The molar ratio of SY02-3B is 19.83:10.34:45.42:9.01:2.23:13.17 with a yield of 2.00%.SY02-3B contains 14 amino acids,among which the content of aspartate,glutamic acid,alanine and glycine is relatively high.We also found that SY02-3 protects palmitic acid-induced INS-1E cell apoptosis,which may be associated with reduced Caspase 3 activity and down-regulated cleaved Caspase 3 protein expression.Conclusion Mulberry leaf polysaccharide SY02-3,which contains acid peptidoglycan(SY02-3A)and proteoglycan(SY02-3B)in mulberry leaves,has the effect of inhibiting PA-induced apoptosis of INS-1E cells.

Mulberry leaf polysaccharideCaspase 3cleaved Caspase 3INS-1E cellsPalimitate

邵赛聪、李茂婷、靳灿、王海颖、丁侃

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上海中医药大学 上海 201203

中国科学院上海药物研究所 上海 201203

中科中山药物创新研究院/中国科学院上海药物研究所 中山 528400

贵州医科大学 贵阳 550025

连云港中医药高等职业技术学校 连云港 222007

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桑叶多糖 Caspase 3活性 cleaved Caspase 3 INS-1E细胞 棕榈酸

国家自然科学基金委员会面上项目上海市"超级博士后"广东省高水平创新研究院项目

3187080120226862021B0909050003

2024

世界科学技术-中医药现代化
中科院科技政策与管理科学研究所,中国高技术产业发展促进会

世界科学技术-中医药现代化

CSTPCD北大核心
影响因子:1.175
ISSN:1674-3849
年,卷(期):2024.26(5)
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