黄芪甲苷对再生障碍性贫血模型小鼠骨髓造血功能及Th17/Treg细胞平衡的影响

Effect of Astragaloside on Bone Marrow Hematopoietic Function and Th17/Treg Cell Balance in Model Mice with Aplastic Anemia

目的:观察黄芪甲苷对免疫介导的再生障碍性贫血模型小鼠骨髓造血功能、相关细胞因子蛋白表达水平及辅助性T淋巴细胞17/调节性T淋巴细胞(Th17/Treg)平衡的影响。方法:采用免疫介导的方法制备再生障碍性贫血小鼠模型。将60只BALB/c小鼠随机分为正常对照组、模型对照组、黄芪甲苷25 mg/kg组、黄芪甲苷50 mg/kg组,每组15只。建模成功后各组灌胃相应药物,每日1次,连续给药28 d。观察小鼠一般情况、体质量、脾脏质量、外周血基本血液学参数;苏木精-伊红(HE)染色法观察骨髓组织病理变化;酶联免疫吸附试验(ELISA)法检测骨髓白细胞介素(IL)-17A、IL-6、转化生长因子-β1(TGF-β1)、IL-10蛋白表达水平;流式细胞术检测黄芪甲苷对小鼠骨髓Th17、Treg细胞比例及Th17/Treg细胞平衡的影响。结果:与模型对照组比较,黄芪甲苷各给药组小鼠一般症状均改善。与正常对照组比较,模型对照组小鼠体质量及脾脏质量下降明显,与模型对照组比较,黄芪甲苷各给药组小鼠体质量、脾脏质量均明显上升(P<0.05)。与模型对照组比较,黄芪甲苷25 mg/kg组白细胞、血小板计数明显升高(P<0.05);黄芪甲苷50 mg/kg组血红蛋白浓度、白细胞、血小板计数明显升高(P<0.05或P<0.01)。黄芪甲苷25 mg/kg组可见造血组织结构修复,造血组织增生度改善;黄芪甲苷50 mg/kg组骨髓组织结构紧密,造血组织增生度明显升高(P<0.05)。黄芪甲苷各给药组骨髓IL-17A、IL-6蛋白表达明显下调,TGF-β1、IL-10蛋白表达明显上调(P<0.05或P<0.01)。流式细胞术检测显示,黄芪甲苷给药组小鼠骨髓Th17细胞比例明显降低,Treg细胞比例明显升高(P<0.05或P<0.01),Th17/Treg细胞比值显著下降(P<0.01)。结论:黄芪甲苷能改善再生障碍性贫血小鼠骨髓造血功能,其机制可能是通过升高Treg细胞比例、降低Th17细胞比例,调节Th17/Treg细胞平衡来实现的。

Objective:To observe the effect of astragaloside on bone marrow hematopoietic function,related cytokine protein expression levels,and Th17/Treg cell balance in immune-mediated aplastic anemia model mice.Methods:An immune-mediated approach was used to prepare model mice of aplastic anemia.There were a total of 60 BALB/c mice,with 15 mice in each group.Mice were randomly divided into a normal control group,a model control group,an astragaloside 25 mg/kg group,and an astragaloside 50 mg/kg group.After successful modeling,each group was orally administered with the corresponding drug once a day for 28 consecutive days.The general condition,body mass,spleen mass,and basic hematological parameters of peripheral blood in mice were observed.HE was used to observe the pathological changes of bone marrow tissue.ELISA was used to detect the protein expression levels of bone marrow IL-17A,IL-6,TGF-β1 and IL-10.Flow cytometry was used to detect the effects of astragaloside on the proportion of Th17 and Treg cells and the balance of Th17/Treg cells in mouse bone marrow.Results:Compared with the model control group,the general symptoms of mice in each treatment group of astragaloside were improved.Compared with the normal control group,the body mass and spleen mass of mice in the model control group decreased significantly,while the body mass and spleen mass of mice in each treatment group of astragaloside increased significantly(P<0.05).Compared with the model control group,the white blood cell count,and platelet count were increased in the astragaloside 25 mg/kg group(P<0.05),and the hemoglobin concentration,white blood cell count,and platelet count were significantly increased in the astragaloside 50 mg/kg group(P<0.05 or P<0.01).The group treated with astragaloside at a dose of 25 mg/kg showed repair of hematopoietic tissue structure and improvement in hematopoietic tissue proliferation.The bone marrow tissue structure was tightly packed and the degree of hematopoietic tissue proliferation was significantly increased in the astragaloside 50 mg/kg group(P<0.05).The expression of bone marrow IL-17A and IL-6 proteins was significantly down-regulated in each group treated with astragaloside glycoside,while the expression of TGF-β1 and IL-10 proteins was significantly up-regulated(P<0.05 or P<0.01).Flow cytometry analysis showed that the proportion of Th17 cells in the bone marrow of mice treated with astragaloside was significantly reduced,while the proportion of Treg cells was significantly increased(P<0.05 or P<0.01),and the Th17/Treg cell ratio was significantly decreased(P<0.01).Conclusion:Astragaloside can improve bone marrow hematopoietic function in mice with aplastic anemia,and its mechanism may be achieved by increasing the proportion of Treg cells,decreasing the proportion of Th17 cells,and regulating the balance of Th17/Treg cells.