| 摘要: |
| 【目的】筛选制备W/O型黄芪多糖（APS）、紫锥菊提取物（EM）纳米乳佐剂的最佳配方，并考察该纳米乳佐剂的免疫效应。【方法】根据伪三元相图中纳米乳区面积的大小并结合肉眼观察，筛选制备黄芪多糖与紫锥菊提取物纳米乳佐剂的配方，观测制剂的形态、粒径分布、pH、黏度及稳定性；以卵清白蛋白（OVA）为模式抗原，用含不同质量黄芪多糖与紫锥菊提取物的纳米乳佐剂免疫小鼠后，测定并比较血清OVA诱导特异性抗体水平的变化。【结果】制备黄芪多糖与紫锥菊提取物纳米乳的配方中，Tween 80、Span 80、石蜡油、乙醇、水的体积比为3∶1∶1.5∶0.5∶\1；所得纳米乳为淡黄色透明液体，透射电镜下呈圆球形，平均粒径45.2 nm，pH 6.71，黏度4.60 s，理化性质较稳定。用黄芪多糖与紫锥菊提取物纳米乳和抗原同时免疫小鼠后，未见任何不良反应，且用黄芪多糖与紫锥菊提取物纳米乳免疫的各试验组小鼠，其血清OVA特异性IgG、IgG1和IgG2a抗体水平均显著高于OVA抗原对照组（P<0.05），而各试验组的IgG、IgG1和IgG2a抗体水平无显著差异；只有APS 200 μg+EM 200 μg+OVA抗原组和APS 400 μg+EM 200 μg+OVA抗原组的IgG、IgG1和IgG2a抗体水平显著高于铝胶（Alum）+OVA抗原组（P<0.05），且以APS 400 μg+EM 200 μg+OVA抗原组的3种抗体水平最高。【结论】黄芪多糖与紫锥菊提取物纳米乳佐剂制备方法简单，稳定性好，能显著增强机体抗体的产生能力，同时可以增强Th1和Th2的免疫应答反应，具有开发佐剂的应用价值。 |
| 关键词: 黄芪多糖 紫锥菊提取物 纳米乳 免疫佐剂 |
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| 基金项目:国家科技支撑计划奶业发展重大关键技术研究与示范项目(2006BAD04A11) |
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| Preparation of W/O Astragalus polysaccharide and Echinacea extract-based nano-emulsion and its amplified immune response |
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| Abstract: |
| 【Objective】The study was to prepare a W/O Astragalus polysaccharede and Echinacea extract-based nano-emulsion and to investigate its amplified immune response.【Method】The formulation of Astragalus polysaccharede and Echinacea extract-based nano-emulsion was optimized by the range of nano-emulsion zone in phase diagrams and the observation of naked eye.Its physicochemical properties were investigated,including morphology,particle size distribution,pH,viscosity and stability.Astragalus polysaccharede and Echinacea extract-based nano-emulsion 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 Astragalus polysaccharede and Echinacea extract-based nano-emulsion formulation consisted of Tween 80,Span 80,paraffin oil,ethanol and water (with the volume ratio of 3∶1∶1.5∶0.5∶1）,which was light yellow fluid.The nano particles were spherical under transmission electron microscopy with an average diameter of 45.2 nm.The viscosity and pH value of Astragalus polysaccharede and Echinacea extract-based nano-emulsion were 4.60 s and 6.71 respectively.The nano-emulsion system showed good stability.When co-administered with OVA,no obvious side effects were observed in the mice immunized with Astragalus polysaccharede and Echinacea Extract-based nano-emulsion.The serum levels of IgG,IgG1 and IgG2a antibodys in all treatment groups of（APS+EM） nano-emulsion immunized mice were significantly increased compared with the groups of OVA （P<0.05）.The serum levels of IgG,IgG1 and IgG2a antibodys were not significantly different between all treatment groups of(APS+EM)nano-emulsion immunized mice （P>0.05）.But the serum levels of IgG,IgG1 and IgG2a antibodys in the group of APS 200 μg+EM 200 μg+OVA and APS 400 μg+EM 200 μg+OVA were significantly increased compared with the groups of Alum+OVA（P<0.05）,and the level of IgG,IgG1 and IgG2a antibodys in the group of APS 400 μg+EM 200 μg+OVA was clearly higher.【Conclusion】The Astragalus polysaccharede and Echinacea extract-based nano-emulsion could amplify the antibody ability and the Th1 and Th2 immune responses,and could be used as the vaccine adjuvant. |
| Key words: Astragalus polysaccharede Echinacea extract nano-emulsion immune adjuvant |
