黄芪多糖APS10-100组分免疫调节活性及其机制

Immunomodulatory activity and mechanism of Astragalus polysaccharide component APS10-100

为探索黄芪多糖APS10-100组分的单糖组成及对小鼠的免疫调节作用,通过柱前衍生化高效液相色谱法分析对比了黄芪多糖APS10-100组分及黄芪粗多糖中的单糖组成。96只ICR小鼠分为对照组、模型组、APS10-100及黄芪粗多糖低、中、高剂量给药组(50,100,200 mg/kg)等8组,除对照组外,其余各组小鼠腹腔注射环磷酰胺(CTX)制备免疫抑制小鼠,给药组每日灌胃给药1次,共给药30 d。给药结束后测小鼠体质量,采血后脱颈处死各组小鼠,取肝脏、脾脏及胸腺等脏器测质量并计算脏器系数。采用ELISA法检测各组小鼠外周血中IFN-γ、IgG含量及肝组织中抗氧化指标超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px),采用流式细胞术检测小鼠血液中T细胞分型。结果显示,黄芪多糖APS10-100组分单糖组成主要为GalA、Glc、Gal及Ara,物质的量的比率为0.43∶1.0∶0.16∶0.39,与黄芪粗多糖有所不同。与对照组比,高剂量APS10-100与黄芪粗多糖均能抵抗CTX所致的体质量下降(P>0.05)。与模型组比,高剂量APS10-100可极显著降低肝脏及脾脏指数、肝组织MDA含量(P<0.01),极显著升高外周血中IFN-γ、CD3^(+)CD4^(+)、CD3^(+)CD8^(+)、CD4^(+)/CD8^(+)及肝组织GSH-Px活性(P<0.01),显著升高外周血中IgG水平(P<0.05);高剂量黄芪粗多糖能显著降低肝脏指数(P<0.05),显著升高IFN-γ、CD3^(+)CD4^(+)及CD4^(+)/CD8^(+)(P<0.05)。结果表明,黄芪多糖APS10-100组分具有良好抗氧化活性,对CTX所致小鼠免疫抑制具有保护作用,其效果优于黄芪粗多糖。

This study aims to investigate the monosaccharide composition of APS10-100 components of Astragalus polysaccharide,and explore its effects and mechanism on immunosuppression.The monosaccharide composition of APS10-100 and crude polysaccharide were analyzed and compared by high performance liquid chromatography with pre-column derivation.Ninety-six ICR mice were randomly divided into eight groups(n=12).The normal control group,cyclophosphamide model group,ASP10-100 groups(50,100,200 mg/kg)and crude polysaccharide groups(50,100,200 mg/kg).Mice were injected intraperitoneally with cyclophosphamide to establish immunosuppression model except the normal control group.Each dose group was administered once daily by gavage for 30 d.The body weight,liver index,spleen index,thymus index,IFN-γ,IgG and T-cell typing in peripheral blood and antioxidant indexes in liver tissue(SOD,MDA,CAT,GSH-Px)of mice were measured.After 30 d of administration,mice of each group were weighed and killed.Liver,spleen and thymus were weighed to calculate the organ coefficients.IFN-γand IgG contents in peripheral blood and antioxidant indexes(SOD,MDA,CAT,GSH-Px)in liver were detected by ELISA,and T cell types in blood were detected by flow cytometry.The results showed that APS10-100 were comprised of GalA,Glc,Gal and Ara,at a molar ratio of 0.43∶1.0∶0.16∶0.39,which were different from those of crude polysaccharide.Compared with the control group,high dose APS10-100 and crude polysaccharide could resist the body mass decrease caused by cyclophosphamide(P>0.05).Compared with the model group,the high dose APS10-100 group showed significant improvements in liver index,spleen index,MDA,GSH-Px,IFN-γ,CD3^(+)CD4^(+),CD3^(+)CD8^(+),CD4^(+)/CD8^(+)(P<0.01)and IgG(P<0.05).Whereas,high dose of crude polysaccharide significantly improved the liver index,IFN-γ,CD3^(+)CD4^(+)and CD4^(+)/CD8^(+)(P<0.05).These results indicated that APS10-100 has good antioxidant activity and protective effect against cyclophosphamide-induced immunosuppression in mice.Its effect was superior to that of Astragalus crude polysaccharide.