3种常见壳寡糖盐的生物活性比较
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摘要
【目的】比较壳寡糖乳酸盐(COS-LA)、壳寡糖乙酸盐(COS-HAc)及壳寡糖盐酸盐(COS-HCl)的组分及其体外生物活性,为壳寡糖的产业化开发提供参考。【方法】以壳聚糖为原料,使用不同酸溶解,在相同酶降解条件下分别制备COS-LA、COS-HAc、COS-HCl,采用超高压液相色谱-飞行时间质谱仪(UPLC-Q TOF MS)分析其组分。以小鼠巨噬细胞RAW264.7为例,研究了3种壳寡糖盐及其对应钠盐对免疫细胞的诱导活性及抑制炎症活性;以白色念珠菌为例,研究了3种壳寡糖盐及其对应钠盐的生物被膜破坏活性。【结果】超高压液相色谱-飞行时间质谱仪分析结果表明,同一聚合度的3种壳寡糖盐的相对含量无明显差异。同等质量浓度下,3种壳寡糖盐的体外免疫诱导活性表现为:COS-LA>COS-HAc>COS-HCl;抗炎活性表现为:COS-LA>COS-HAc>COS-HCl;生物被膜破坏活性表现为:COS-LA>COS-HAc>COS-HCl。3种壳寡糖盐对应钠盐的生物活性检测结果与壳寡糖盐结果相似。【结论】盐型对壳寡糖的生物活性有一定的影响,其中以壳寡糖乳酸盐的生物活性最佳。
【Objective】This study compared the composition and bioactivity of chitosan oligosaccharide hydrolactate(COS-LA),chitosan oligosaccharide acetate acid(COS-HAc)and chitosan oligosaccharide hydrochloride(COS-HCl)to provide reference for industrialization development of chitosan oligosaccharides.【Method】COS-LA,COS-HA and COS-HCl were prepared by enzymatic degradation with chitosan using different acids.The components were analyzed by UPLC-Q TOF MS.The induction and inhibition of inflammatory activity of three COS salts and their corresponding sodium salts were researched using RAW264.7 cells and the antibacterial activity was studied with candida albicans.And the result of bioactivity of sodium salts is similar with COS salts.【Result】The results of UPLC-Q TOF MS showed that there was no significant difference in the composition of chitosan oligosaccharide salts with the same polymerization degree.At same concentration,immunogenic activity was in the order of COS-LA>COS-HAc>COSHCl.Anti-inflammatory activity was in the order of COS-LA>COS-HAc>COS-HCl and antibacterial activity was in the order of COS-LA>COS-HAc>COS-HCl.【Conclusion】The bioactivity of chitosan oligosaccharide salts was influenced by salt type.Chitosan oligosaccharide hydrolactate(COS-LA)had the best bioactivity.
【Objective】This study compared the composition and bioactivity of chitosan oligosaccharide hydrolactate(COS-LA),chitosan oligosaccharide acetate acid(COS-HAc)and chitosan oligosaccharide hydrochloride(COS-HCl)to provide reference for industrialization development of chitosan oligosaccharides.【Method】COS-LA,COS-HA and COS-HCl were prepared by enzymatic degradation with chitosan using different acids.The components were analyzed by UPLC-Q TOF MS.The induction and inhibition of inflammatory activity of three COS salts and their corresponding sodium salts were researched using RAW264.7 cells and the antibacterial activity was studied with candida albicans.And the result of bioactivity of sodium salts is similar with COS salts.【Result】The results of UPLC-Q TOF MS showed that there was no significant difference in the composition of chitosan oligosaccharide salts with the same polymerization degree.At same concentration,immunogenic activity was in the order of COS-LA>COS-HAc>COSHCl.Anti-inflammatory activity was in the order of COS-LA>COS-HAc>COS-HCl and antibacterial activity was in the order of COS-LA>COS-HAc>COS-HCl.【Conclusion】The bioactivity of chitosan oligosaccharide salts was influenced by salt type.Chitosan oligosaccharide hydrolactate(COS-LA)had the best bioactivity.
引文
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[3]Lodhi G,Kim Y S,Hwang J W,et al.Chitooligosaccharide and its derivatives:preparation and biological applications[J].Biomed Research International,2014,2014:doi:10.1155/2014/654913.
[4]Kerch G.The potential of chitosan and its derivatives in prevention and treatment of age-related diseases[J].Marine Drugs,2015,13(4):2158-2182.
[5]Li K C,Xing R,Liu S,et al.Advances in preparation,analysis and biological activities of single chitooligosaccharides[J].Carbohydrate Polymers,2016,139:178-190.
[6]李昱,许青松,彭强,等.壳寡糖抗炎作用研究进展[J].中国生化药物杂志,2012,33(3):326-328.Li Y,Xu Q S,Peng Q,et al.Research progress on anti-inflammatory effects of chitooligosaccharides[J].Chinese Biochemical Pharmaceutics,2012,33(3):326-328.
[7]柯海萍,赵鲁杭.壳寡糖对体外培养的巨噬细胞IL-18基因表达的影响[J].海军医学杂志,2004,25(2):97-99.Ke H P,Zhao L H.Effects of oligochitosan on IL-18gene expression in cultured macrophages[J].Navy Medicine,2004,25(2):97-99.
[8]Yoon H J,Moon M E,Park H S,et al.Chitosan oligosaccharide(COS)inhibits LPS-induced inflammatory effects in RAW264.7macrophage cells[J].Biochemical&Biophysical Research Communications,2007,358(3):954-959.
[9]Yang E J,Kim J G,Kim J Y,et al.Anti-inflammatory effect of chitosan oligosaccharides in RAW264.7cells[J].Open Life Sciences,2010,5(1):95-102.
[10]Qiao Y,Ruan Y Y,Xiong C N,et al.Chitosan oligosaccharides suppressant LPS binding to TLR4/MD-2receptor complex[J].Carbohydrate Polymers,2010,82(2):405-411.
[11]Liu B,Wang X Y,Pang C S,et al.Preparation and antimicrobial property of chitosan oligosaccharide derivative/rectorite nanocomposite[J].Carbohydrate Polymers,2013,92(2):1078.
[12]Zou P,Yang X,Wang J,et al.Advances in characterisation and biological activities of chitosan and chitosan oligosaccharides[J].Food Chemistry,2016,190:1174-1181.
[13]Schimpl M,Rush C L,Betou M,et al.Human YKL-39is a pseudo-chitinase with retained chitooligosaccharide-binding properties[J].Biochemical,2012,446(1):149-157.
[14]Younes I,Rinaudo M.Chitin and chitosan preparation from marine sources.structure,properties and applications[J].Marine Drugs,2015,13(3):1133.
[15]Jung W J,Park R D.Bioproduction of chitooligosaccharides:present and perspectives[J].Marine Drugs,2014,12(11):5328.
[16]Carre1o-gómez B,Duncan R.Evaluation of the biological properties of soluble chitosan and chitosan microspheres[J].Pharmaceutics,1997,148(2):231-240.
[17]Woraphatphadung T,Kowapradit J,Ngawhirunpat T,et al.Effect of salt forms of chitosan on in vitro permeability enhancement in intestinal epithelial cells(Caco-2)[J].Pharmaceutical Research,2013,12(4):495-501.
[18]Huanbutta K,Cheewatanakornkool K,Terada K,et al.Impact of salt form and molecular weight of chitosan on swelling and drug release from chitosan matrix tablets[J].Carbohydrate Polymers,2013,97(1):26-33.
[19]Zhang H,Du Y G,Yu X J,et al.Preparation of chitooligosaccharides from chitosan by a complex enzyme[J].Carbohydrate Research,1999,320(3/4):257-260.
[20]Gerlier D,Thomasset N.Use of MTT colorimetric assay to measure cell activation[J].Journal of Immunological Methods,1986,94(1):57-63.
[21]李恩民,魏大愚,许丽艳,等.间接测定生物组织中一氧化氮含量的Griess法的改良[J].汕头大学医学院学报,1999(1):75-77.Li E M,Wei D Y,Xu L Y,et al.An improved Griess method for determining the content of nitric oxide in biological tissues[J].Journal of Shantou University Medical College,1999(1):75-77.