高压灭菌前后党参口服液中多糖的免疫活性研究
Study of immunocompetence of polysaccharides from Codonopsis pilosula oral solution before and after autoclaving
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摘要: 研究高压灭菌前后党参口服液中多糖的免疫活性变化。采用醇沉法、超滤膜分离法得到党参口服液中活性多糖部分,以RAW264.7巨噬细胞为模型,通过对细胞增殖指数、吞噬指数、NO释放量以及细胞因子(IL-6、IL-1β、TNF-α)的分泌量的测定,评价高压灭菌前后党参口服液中多糖的免疫活性。结果表明:超滤分离得到高压灭菌前以及高压灭菌后党参口服液中分子量小于30 k Da的多糖组分CPP-2和HCPP-2,其得率分别为22.41%和15.91%,纯度分别为69.03%和69.30%,且二者均能较好地促进巨噬细胞增殖(增殖指数=1.131.48),因此CPP-2和HCPP-2是党参口服液中主要的活性多糖部分。与CPP-2相比,HCPP-2在25400μg/m L浓度范围内作用于巨噬细胞的吞噬能力、TNF-α和IL-1β生成显著性降低(p<0.05);在100400μg/m L浓度范围内,NO和IL-6生成显著性降低(p<0.05)。说明高压灭菌会导致党参口服液中多糖的免疫活性降低。Abstract: To study the changes of immunocompetence of polysaccharides from Codonopsis pilosula oral solution before and after autoclaving, the methods of alcohol precipitation and ultrafiltration membrane were used, the active polysaccharides of Codonopsis pilosula oral solution before and after autoclaving were obtained, and their immune activities were evaluated by measuring inflammatory responses in RAW264.7 macrophages of cell proliferation index, phagocytic index, NO release and secretion of cytokines (IL-6, IL-1β, TNF-α) .The results showed that the yields of polysaccharide fraction with molecular weight less than 30 k Da before autoclaving and autoclaving in Codonopsis pilosula oral solution (COP-2 and HCOP-2) were22.41% and 15.91%, the purity were 69.03% and 69.30%, respectively, and both of them could promote the proliferation of macrophages (proliferative index = 1.13~1.48) , which means they were the main active polysaccharidesin Codonopsis pilosula oral solution.Compared with COP-2, HCOP-2 decreased the phagocytic capacity, TNF-α and IL-1β production in the concentration range of 25~400 μg/m L (p < 0.05) , NO and IL-6 production were significantly decreased in the concentration range of 100~400 μg/m L (p < 0.05) , which indicated that autoclave resulted in decrease of immune activity of Codonopsis pilosula oral solution active polysaccharide.