黄芪多糖对系统性红斑狼疮小鼠免疫调节的影响

Effect of astragalus polysaccharide on immune regulation of systemic lupus erythematosus mice

目的探讨黄芪多糖对系统性红斑狼疮MRL/lpr小鼠免疫功能的影响及作用机制。方法将20只MRL/lpr狼疮小鼠依据随机数字表法分为模型组和药物组,每组各10只,另选择10只雌性C57BL/6小鼠作为对照组。药物组采用黄芪多糖治疗,对照组和模型组均采用同等剂量生理盐水治疗。比较各组小鼠的自身抗体水平。采用流式技术法测定各组脾脏组织辅助性T细胞1(Th1)和辅助性T细胞2(Th2)细胞比例,使用蛋白质印迹法(Western blotting)和q-PCR技术测定T盒转录因子(T-bet)和GATA结合蛋白3(GATA-3)的表达情况。本次研究起止时间为2019年2—6月。结果模型组小鼠血清抗核抗体(ANA)、抗双链脱氧核糖核酸抗体(Anti-dsDNA)和抗小核糖核蛋白/Sm抗体(Anti-snRNP/Sm)水平分别为(38.11±3.84)ng/L、(524.63±52.69)μmol/L、(23.05±2.31)ng/L,与对照组(24.23±2.41)ng/L、(340.26±34.37)μmol/L、(15.01±1.51)ng/L相比均明显升高(P<0.05);药物组小鼠血清ANA抗体、Anti-dsDNA抗体和Anti-snRNP/Sm抗体水平分别为(29.45±2.95)ng/L、(468.14±46.88)μmol/L、(18.44±1.83)ng/L,与模型组相比明显降低(P<0.05)。模型组小鼠Th1亚群比例、Th2亚群比例分别为(11.32±1.13)%、(2.84±0.29)%,与对照组小鼠(18.95±1.71)%、(2.21±0.22)%相比,模型组小鼠Th1亚群比例明显降低,Th2亚群比例明显升高(P<0.05);与模型组相比,药物组小鼠Th1亚群比例(14.69±1.47)%明显升高,Th2亚群比例(2.49±0.24)%明显降低(P<0.05)。与对照组小鼠比较,模型组小鼠T-bet蛋白表达和mRNA水平显降低,GATA-3蛋白和mRNA表达水平均显著升高(P<0.05);与模型组小鼠相比,药物组小鼠T-bet蛋白和mRNA表达水平明显升高,GATA-3蛋白和mRNA表达水平显著降低(P<0.05)。结论黄芪多糖可影响MRL/lpr狼疮小鼠T-bet、GATA3的表达水平,调节Th1/Th2细胞的平衡,进而下调自身抗体水平发挥治疗作用。

Objective To investigate the effect and mechanism of Astragalus polysaccharide on immune function of MRL/lpr lupus mice.Methods Twenty MRL/lpr lupus mice were randomly assigned into model group and drug group according to the random number table method,with 10 mice in each group,and 10 female C57BL/6 mice were selected as control group.The drug group was treated with Astragalus polysaccharide,and the control group and the model group were treated with the same dose of normal saline.The autoantibody levels of each group of mice were compared.The ratio of helper T cell 1(Th1)and helper T cell 2(Th2)cells in spleen tissues of each group was determined by flow cytometry.The expression of T-box expressed in T cells(T-bet)and GATA binding protein 3(GATA-3)was determined by Western blotting and q-PCR.The starting and ending time of this study was from February 2019 to June 2019.Results The serum levels of antinuclear antibody(ANA),anti double stranded DNA antibody(anti-dsDNA)and anti small nuclear ribonucleoprotein/Sm antibody(anti-snRNP/Sm)in model group were(38.11±3.84)ng/L,(524.63±52.69)μmol/L and(23.05±2.31)ng/L,respectively,which were significantly higher than those in the control group[(24.23±2.41)ng/L,(340.26±34.37)μmol/L,(15.01±1.51)ng/L,P<0.05].The serum levels of ANA antibody,anti-dsDNA antibody and anti-snRNP/Sm antibody in the drug group were(29.45±2.95)ng/L,(468.14±46.88)μmol/L and(18.44±1.83)ng/L,respectively,which were significantly lower than those in the model group(P<0.05).The proportion of Th1 and Th2 subgroups in the model group were(11.32±1.13)%and(2.84±0.29)%,and in the control group were(18.95±1.71)%and(2.21±0.22)%.The proportion of Th1 subgroup in the model group was significantly decreased,while the proportion of Th2 subgroup was significantly increased(P<0.05).Compared with the model group,the proportion of Th1 subgroup in the drug group(14.69±1.47)%was increased,while the proportion of Th2 subgroup(2.49±0.24)%was decreased(P<0.05).Compared with the control group,the T-bet protein expression and mRNA levels in the model group and the drug group were significantly decreased,and the GATA-3 protein and mRNA expression levels were significantly increased(P<0.05).Compared with the model group,the expression levels of T-bet protein and mRNA in the drug group were significantly increased,and the expression levels of GATA-3 protein and mRNA were significantly decreased(P<0.05).Conclusion Astragalus polysaccharide can affect the expression level of T-bet and GATA-3 in MRL/lpr lupus mice,regulate the balance of Th1/Th2 cells,and down-regulate the level of autoantibodies.