GAO Zhenzhen,ZHANG Chao,JING Lirong,et al.The comparison of seven selenizing polysaccarides in vitro and in vivo immune-enhancing activity in mouse[J].Journal of Nanjing Agricultural University,2020,43(4):705-713.[doi:10.7685/jnau.201908044]





The comparison of seven selenizing polysaccarides in vitro and in vivo immune-enhancing activity in mouse
高珍珍1 张超1 景丽荣1 胡元亮2
1. 内蒙古农业大学兽医学院, 内蒙古 呼和浩特 010018;
2. 南京农业大学动物医学院, 江苏 南京 210095
GAO Zhenzhen1 ZHANG Chao1 JING Lirong1 HU Yuanliang2
1. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
selenizing polysaccharidelymphocyteproliferationimmunoglobulincytokinesmouse
[目的] 本试验旨在筛选出增强免疫活性最强的硒化多糖,为研制硒多糖类免疫增强剂提供理论依据。[方法] 体外试验:测定硒化当归多糖(sCAPS)、硒化山药多糖(sDOP)、硒化党参多糖(sCPPS)、硒化淫羊藿多糖(sEPS)、硒化大蒜多糖(sGPS)、硒化百合多糖(sLP)、硒化白术多糖(sAMP)7种硒化多糖(sPS)和修饰试剂亚硒酸钠(Na2SeO3)对小鼠脾脏淋巴细胞的安全浓度,取安全范围内5种浓度的各硒化多糖和Na2SeO3单独或与PHA、LPS同时加入小鼠脾脏的淋巴细胞培养体系,测定淋巴细胞增殖率,筛选出增强免疫效果较好的3种硒化多糖sCAPS、sCPPS和sGPS。体内试验:测定各组小鼠免疫卵清蛋白(OVA)时,分别灌胃150、100、50 μg·mL-1的sCAPS、sCPPS、sGPS和10 μg·mL-1 Na2SeO3各0.4 mL,免疫对照组和空白对照组灌胃等量生理盐水,每天1次,连续3 d,分别于首免后7、21和35 d每组随机抽取8只小鼠摘眼球采血,分离血清,用ELISA试剂盒测定血清IgG、IgM、IL-2、IL-4和IFN-γ含量,HE染色后比较各组小鼠脾脏的组织学变化。[结果] 在体外,单独加入或与PHA、LPS同时加入时,sCAPS、sCPPS和sGPS组的淋巴细胞增殖率均高于或显著高于其他各组(P<0.05);在小鼠体内,100 μg·mL-1 sCPPS处理时,免疫球蛋白和细胞因子含量多高于或显著高于其他各组(P<0.05),脾脏白髓和边缘区面积增多最明显。[结论] sCPPS、sCAPS和sGPS的体外增强免疫活性较强,100 μg·mL-1sCPPS处理小鼠的体内增强免疫活性最强,可作为硒多糖类免疫增强剂的候选。
[Objectives] This study was conducted to screen out the best selenizing polysaccharide with immune-enhancing activities,and provide the materials for developing immunopotentiators.[Methods] Seven selenizing polysaccarides(sPS),selenizing Chinese Angelica polysaccharide(sCAPS),selenizing Dioscorea oppositae thunb polysaccharides(sDOP),selenizing Codonopsis pilosula polysaccharide(sCPPS),selenizing Epimedium polysaccharide(sEPS),selenizing garlic polysaccharide(sGPS),selenizing lily polysaccharide(sLP),selenizing Atractylodes macrocephala polysaccharide(sAMP) and sodium selenite Na2SeO3 diluted with RPMI1640 at five concentrations were added into the cultured mouse spleen lymphocytes in single stimulation of sPS,stimulation of sPS with PHA or LPS,respectively. The changes of the lymphocyte proliferation rate were determined and compared,three selenizing polysaccharides were screened out from in vitro test,at three concentrations of 150,100,50 μg·mL-1,and Na2SeO3 at 10 μg·mL-1 with 0.4 mL were further compared on immune-enhancing activities in vivo,and the changes of serum IgG,IgM,IL-2,IL-4,IFN-γ and the histological changes of the spleen were determined and compared.[Results] The results showed that in single stimulation,synergistical stimulation with PHA or LPS,the lymphocyte proliferation rates of sCAPS,sCPPS and sGPS groups were higher or significantly higher than the other groups(P<0.05). The immunoglobulin and cytokines contents of selenizing polysaccharide groups were significantly higher than those of corresponding immunization control group,and the 100 μg·mL-1 sCPPS at the most time points were higher or significantly higher than the other groups(P<0.05). The growth center of the white pulp(WP) and the lymphocytes in the margin zone(MZ) in 100 μg·mL-1 sCPPS significantly increased.[Conclusions] These results indicated that,in vitro test,the actions of sCPPS,sCAPS and sGPS were stronger,the 100 μg·mL-1 sCPPS group possessed the strongest effects and in vivo test,and sCPPS could be used as a drug candidate for selenizing polysaccharide immunopotentiators,and provide references for clinical application.


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更新日期/Last Update: 1900-01-01