Extraction,purification,characterization and anti-inflammation of glyco-protein from Codonopsis pilosula Nannf.var.modesta(Nannf.)L.T.Shen
刘子琴 1王颖莉 2赵颖 1李文汇 1宋亮亮 1王靖 1王宏月 1杜红1
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作者信息
1. 北京中医药大学,北京 102488
2. 山西中医药大学,太原 030619
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摘要
目的 对党参中的糖蛋白(即党参总糖蛋白,CGP)进行分离、纯化、结构表征和抗炎药效研究.方法 使用Tris-HCL缓冲溶液在39 ℃水浴提取CGP,采用Q-Sepharose Fast Flow强阴离子交换层析和SDS-PAGE法对CGP进行分离纯化.使用紫外光谱、圆二色谱等方法,对纯品组分的结构及组成进行研究.以脂多糖(LPS)诱导的RAW264.7为炎症模型,验证CGP的抗炎药效.结果 得到分子量分别为35 kDa和25kDa的两种纯品组分CGP1和CGP2,其结构均为O-糖肽键连接的糖蛋白,并证明5mg·mL-1的CGP抗炎效与槲皮素相当.结论 CGP有望成为一种具有抗炎功效的潜在物质成分,为党参的药食同源应用提供了科研依据.
Abstract
Objective To extract,purifying,characterize the glycoprotein(CGP)in Codonopsis pilosula Nannf.var.modesta(Nannf.)L.T.Shen,and to determine its anti-inflammation effect.Methods CGP was extracted by water bath at 39 ℃ with Tris-HCL buffer solution,followed by purification through the Q-Sepharose anion exchange chromatography and SDS-PAGE.Structural characterization of the pure fractions was evaluated by UV spectroscopy and circular dichroism and other analytical techniques.The inhibition of CGP on the cell inflammation factors was observed with LPS-stimulated RAW 264.7 as the inflammation model.Results Two pure CGP fractions with molecular weights of 35 kDa and 25 kDa were obtained,both structured as glycoprotein linked by O-glycosyl peptide bonds.The experiments demonstrated that the anti-inflammation efficacy of CGP at 5 mg·mL-1 was equivalent to that of quercetin.Conclusion CGP holds promise as a potential component with anti-inflammation effect,providing scientific basis for the homology of medicine and food of Codonopsis pilosula Nannf.var.modesta(Nannf.)L.T.Shen.
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