响应曲面法优化党参多糖PLGA纳米颗粒的包封条件及其对RAW264.7的影响  

Optimization of encapsulated conditions of Codonopsis pilosula polysaccharide PLGA nanoparticles using response surface methodology and their effects on RAW264.7

作　　者：王涛 伍智天 许逸舟 郭劲航 任喆 秦韬 WANG Tao;WU Zhitian;XU Yizhou;GUO Jinhang;REN Zhe;QIN Tao(Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Heath in Fujian Province/College of Animal Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学动物科学学院福建省兽医中药与动物保健重点实验室,福建福州350002

出　　处：《中国兽医科学》2024年第9期1267-1276,共10页Chinese Veterinary Science

基　　金：福建省自然科学基金面上项目(2023J01074);福建农林大学科技创新专项基金项目(KFB23102)。

摘　　要：本试验旨在探寻聚乳酸-羟基乙酸共聚物(poly lactic-co-glycolic acid,PLGA)加载对党参多糖(Codonopsis pilosula polysaccharide,CPP)结构特征和免疫调控作用的影响。采用乳化溶剂挥发法制备党参多糖PLGA纳米颗粒(CPP-PLGA-NPs),应用响应曲面法优化包封条件,通过透射电镜、Zeta电位与纳米粒度仪分析其结构,并利用MTT法、流式细胞术、ELISA法测定CPP-PLGA-NPs和CPP对RAW264.7细胞的免疫调控作用。结果显示,制得的纳米颗粒表面光滑、形态均匀、分散性良好、状态稳定、粒径小。最佳制备条件为:有机相与内水相之比为8.51,外水相与初乳比为7.12,F68的浓度为0.33%,PLGA的浓度为30 mg/m L,制备的纳米颗粒的包封率为93.54%±0.56%。与空白组相比,CPP-PLGA-NPs和CPP都显著提高了巨噬细胞的吞噬活性、NO分泌量、CD40和CD86的表达量、细胞因子(TNF-α、IL-6、IL-1β)的分泌量(P<0.05),在浓度为0.97μg/m L时CPP-PLGA-NPs调控作用达到最高。结果表明,CPP-PLGA-NPs和CPP都有较好的免疫调节活性,为党参多糖的开发与利用提供了理论依据。The aim of this experiment was to investigate the effects of poly lactic-co-glycolic acid(PLGA)loading on the structural characteristics and immune regulation of Codonopsis pilosula polysaccharide(CPP).Codonopsis pilosula polysaccharide PLGA nanoparticles(CPP-PLGA-NPs)were prepared by emulsified solvent volatilization method.The encapsulated conditions were optimized by Response surface method.The structures were analyzed by transmission electron microscopy,Zeta potential and nanoparticle size analyzer.The immunoregulatory effects of CPP-PLGA-NPS and CPP on RAW264.7 cells were determined by MTT,flow cytometry and ELISA.The results show that the prepared nanoparticles have smooth surface,uniform morphology,good dispersion,stable state and small particle size.The optimum preparation conditions were as follows:the ratio of organic phase to inner water phase was 8.51,the ratio of outer water phase to colostrum was 7.12,the concentration of F68 was 0.33%,the concentration of PLGA was 30 mg/mL,and the encapsulation rate of prepared nanoparticles was 93.54%±0.56%.Compared with blank group,CPP-PLGA-NPS and CPP significantly(P<0.05)increased phagocytosis activity of macrophages,secretion of NO,expression of CD40 and CD86,secretion of cytokines(TNF-α,IL-6,IL-1β).The regulatory effect of CPP-PLGA-NPs reached the highest at 0.97μg/mL.The results showed that both CPP-PLGA-NPS and CPP had good immunomodulatory activity,which provided a theoretical basis for the development and utilization of Codonopsis pilosula polysaccharide.

关 键 词：党参多糖 PLGA 响应曲面法 免疫调控 

分 类 号：S853.74[农业科学—临床兽医学]