中国中药杂志

2010, v.35(04) 439-443

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人参皂苷纳米乳的制备及其免疫佐剂效应研究
Preparation of O/W ginseng saponins-based nanoemulsion and its amplified immune response

曹发昊;欧阳五庆;王艳萍;
CAO Fahao1,OUYANG Wuqing 1,WANG Yanping2(1.College of Veterinary Medicine,Northwest Agricultural and Forestry University,Yangling 712100,China;2.Shangqiu Vocational and Technical College,Shangqiu 476000,China)

摘要(Abstract):

目的:筛选最佳处方制备O/W人参皂苷纳米乳,考察其免疫佐剂效应。方法:根据纳米乳区和人参皂苷的溶解性,筛选人参皂苷纳米乳的处方;测定制剂的形态、粒径分布、pH、黏度及稳定性;以OVA为模式抗原,测定人参皂苷纳米乳免疫小鼠后血清OVA诱导特异性抗体水平。结果:筛选出Cremophor RH40/甘油/IPM/水(Km=1∶1)制备20 g.L-1人参皂苷纳米乳,为淡黄色透明液体,透射电镜下呈圆球形,平均粒径为72.2 nm,pH6.02,黏度4.20 s,理化性质较稳定;人参皂苷纳米乳和抗原同时免疫小鼠后,未见任何不良反应,GS-NE组IgG,IgG1和IgG2a抗体水平均显著高于OVA组和GS组;GS-NE组的IgG和IgG1抗体水平与铝佐剂组无显著性差异,但GS-NE组IgG2a抗体水平均显著高于铝佐剂组。结论:人参皂苷纳米乳稳定性很好,能增强机体对抗原的抗体产生能力,同时增强Th1和Th2免疫应答反应,具有开发佐剂的应用价值。
Objective: To prepare an O/W ginseng saponins-based nanoemulsion and investigate its amplified immune response.Method: The formulation of ginseng saponins-based nanoemulsion was optimized via the range of nanoemulsion zone in phase diagrams and the solubility of ginseng saponins.Its physicochemical properties were investigated,including morphology,particle size distribution,pH,viscosity and stability.Ginseng saponins-based nanoemulsion as adjuvant was co-administrated with a model antigen ovalbumin(OVA)in mice.Two weeks after the boosting,the serum levels of OVA-specific antibody and its isotypes were determined.Result: The optimized ginseng saponins-based nanoemulsion formulation consisted of ginseng saponins,IPM,Cremophor RH 40,glycerol and water(with the weight ratio of 2∶4∶17.8∶17.8∶58.4),which was a light yellow fluid.The shape of droplets was spherical under transmission electron microscopy with an average diameter of 72.20 nm and a polydispersity index of 0.052.The viscosity and pH value of it were 4.20s and 6.02,respectively.And it showed good stability.When co-administered with OVA,no obvious side effects were observed in the mice immunized with ginseng saponin-based nanoemulsion.The serum levels of IgG,IgG1 and IgG2a antibody in the group of ginseng saponin-based nanoemulsion immunized mice was significantly increased compared to the groups of OVA and the saline solution of ginseng saponin.Compared with the adjuvant aluminium hydroxide,the serum levels of IgG and IgG1 antibodys in the groups of ginseng saponins-based nanoemulsion had no significant difference,but the level of IgG2a was obviously higher.Conclusion: ginseng saponin-based nanoemulsion could amplify the Th1 and Th2 immune responses,and can be used as the vaccine adjuvant.

关键词(KeyWords): 人参皂苷;纳米乳;佐剂;铝胶
ginseng saponinss;nanoemulsion;adjuvant;aluminium hydroxide

Abstract:

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基金项目(Foundation): 国家科技支撑计划项目(2006BAD04A11)

作者(Author): 曹发昊;欧阳五庆;王艳萍;
CAO Fahao1,OUYANG Wuqing 1,WANG Yanping2(1.College of Veterinary Medicine,Northwest Agricultural and Forestry University,Yangling 712100,China;2.Shangqiu Vocational and Technical College,Shangqiu 476000,China)

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