穿龙薯蓣皂苷元对CIA小鼠脾脏Treg细胞的影响

Effects of Diosgenin on Treg Cell in Spleen of CIA Mice

目的:在胶原性关节炎小鼠模型中初步探讨穿龙薯蓣皂苷元对其脾脏Treg细胞的影响。方法:通过将鸡Ⅱ型胶原乳剂与弗氏完全佐剂混合后免疫DBA1/J小鼠,建立胶原性关节炎(CIA)模型。通过免疫磁珠分离法(MACS)分选CIA小鼠脾脏中的CD4+CD25+Treg细胞。流式细胞术检测Treg细胞的纯度,并体外培养。穿龙薯蓣皂苷元作用48h后,流式细胞术检测Treg细胞比例,实时荧光定量PCR法检测其转录因子Foxp3的表达,ELISA法检测相关细胞因子IL-10的水平。结果:成功建立CIA模型小鼠。MACS法分选CIA小鼠脾脏Treg细胞纯度达90%以上。穿龙薯蓣皂苷元作用48h后,与对照组相比,给药组Treg细胞比例显著升高(P<0.05),Foxp3 mRNA表达显著升高(P<0.05),细胞上清中IL-10的水平显著升高(P<0.05)。结论:穿龙薯蓣皂苷元能显著提高CIA小鼠脾脏中Treg细胞的比例、转录因子Foxp3和相关细胞因子IL-10的水平。

Objective:To evaluate the effect of diosgenin on regulatory Treg cells in spleen of collagen induced arthritis(CIA)mice.Methods:The CIA mice model was established by immunizing DBA1/J mice with chicken type II collagen and Freund’s completed adjuvant.And CD4+CD25+Treg cells in spleen of CIA model mice were isolated by MACS.Then,to detect the purity of Treg cells by flow cytometry and culture them in vitro.48 hours after treatment,the proportion of Treg cells was detected by flow cytometry,the expression of transcription factor Foxp3 in Treg cells was detected by real-time fluorescence quantitative PCR,and the levels of IL-10 were detected by ELISA.Results:CIA model mice were established successfully.The purity of Treg cells isolated by MACS was more than 90%.Compared with the control group,the percentage of Treg cells,the Foxp3 mRNA level,and the expression of IL-10 in cell supernatant were increased significantly(P<0.05)48 hours after treatment.Conclusion:Diosgenin can significantly increase the proportion of Treg cells,the levels of transcription factor Foxp3 and IL-10 in CIA mice.