黄芪多糖与紫锥菊提取物纳米乳的制备及免疫佐剂效应研究  

Preparation of W/O Astragalus polysaccharide and Echinacea extract-based nano-emulsion and its amplified immune response

作　　者：李树鹏[1,2] 郝艳霜[2] 欧阳五庆[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]河北农业大学中兽医学院,河北定州073000

出　　处：《西北农林科技大学学报（自然科学版）》2011年第10期10-16,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：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)

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45.2nm,pH 6.71,黏度4.60s,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 200μg+EM 200μg+OVA抗原组和APS400μg+EM 200μg+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 400μg+EM 200μg+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。【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（P0.05）.The serum levels of IgG,IgG1 and IgG2a antibodys were not significantly different between all treatment groups of（APS＋EM） nano-emulsion immunized mice（P0.05）.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（P0.05）,and the level of IgG,IgG1 and IgG2

关 键 词：黄芪多糖 紫锥菊提取物 纳米乳 免疫佐剂 

分 类 号：S852.43[农业科学—基础兽医学]