再障膏方对再生障碍性贫血小鼠FOXP3、GATA-3、T-bet、Notch1蛋白表达及Treg/Th17的影响

Effects of Zaizhang cream formula on the protein expression of FOXP3,GATA-3,T-bet,Notch1 and Treg/Th17 in mice with aplastic anemia

目的观察再障膏方联合环孢素对再生障碍性贫血模型小鼠脾脏叉头状转录因子3(forkhead box protein 3,FOXP3)、GATA结合蛋白3(GATA binding protein 3,GATA-3)、T细胞特异性转录因子(transcription factor T-bet,T-bet)、Notch1蛋白表达及外周血调节性T细胞(regulatory T cell,Treg)/辅助性T细胞17(helper T type 17,Th17)的影响。方法使用白消安灌胃制作再生障碍性贫血小鼠模型,造模成功后随机分为空白组、模型组、中药组、西药组、中+西药组。中药组予再障膏方溶液灌胃,西药组予Cs A溶液灌胃,中+西药组予再障膏方+Cs A溶液灌胃,连续给药10天。采用流式细胞术检测小鼠外周血Treg、Th17细胞表达;采用蛋白免疫印迹法检测小鼠脾脏FOXP3、GATA-3、T-bet、Notch1蛋白表达水平。结果再障膏方联合环孢素能明显增加再生障碍性贫血小鼠外周血Treg/Th17及脾脏FOXP3、GATA-3、Notch1蛋白表达、降低T-bet蛋白表达(P<0.05),且疗效优于单纯西药组及中药组(P<0.05)。结论再障膏方联合环孢素治疗能显著改善AA模型小鼠脾脏转录因子FOXP3、GATA-3、T-bet的表达,促进外周血Treg/Th17细胞比例恢复平衡,但是否与Notch1信号通路相关有待进一步研究。

Objective To observe the effects of Zaizhang cream formula combined with cyclosporine on the protein expression of FOXP3,GATA-3,T-bet,and Notch1 in spleen and Treg/Th17 in peripheral blood of aplastic anemia(AA)model mice.Methods The mouse model with aplastic anemia was established by gavage of busulfan.After successful modeling,the mice was divided into blank group,model group,Chinese medicine group,western medicine group,Chinese and western medicine group.The Chinese medicine group was given Zaizhang cream formula by gavage,the western medicine group was given CsA solution by gavage,and the Chinese and western medicine group was given Zaizhang cream formula and CsA solution by gavage,for 10 consecutive days.The expressions of Treg and Th17 cells in peripheral blood of mice were detected by flow cytometry.The protein expression of FOXP3,GATA-3,T-bet and Notch1 in spleen were detected by Western Blot.Results Zaizhang cream formula combined with cyclosporine can significantly increased the expression of Treg/Th17 in peripheral blood and the protein expression of FOXP3,GATA-3 and Notch1 of AA mice,and decreased the protein expression of T-bet(P<0.05).The curative effect was better than that of the western medicine group and the Chinese medicine group(P<0.05).Conclusion Treating AA model mice with therapy of Zaizhang cream formula combined with cyclosporine,can significantly improve the expression of transcription factors FOXP3,GATA-3 and t-bet in spleen and promote the restoration of balance of Treg/Th17 cells ration in peripheral blood.Whether it is related to Notch1 signaling pathway remains to be further studied.