穿龙薯蓣皂苷元对DBA1/J小鼠CD4^(+)CD25^(-)T细胞的影响

Effects of Diosgenin on CD4^(+)CD25^(-)T Cell in Spleen of DBA1/J Mice

目的初步探讨穿龙薯蓣皂苷元对DBA1/J小鼠CD4^(+)CD25^(-)T细胞的影响。方法采用MACS磁珠分选法分离DBA1/J小鼠脾CD4^(+)CD25^(-)T细胞,并鉴定纯度。穿龙薯蓣皂苷元作用CD4^(+)CD25^(-)T细胞后,采用ELISA方法检测各组培养上清中IL-10的浓度,Real-Time PCR方法检测Foxp3的水平,FCM检测Treg细胞的比率。结果免疫磁珠术分选DBA1/J小鼠脾CD4^(+)CD25^(-)T细胞纯度达93.89%。给药作用48h后,与对照组相比,穿龙薯蓣皂苷元组上清中IL-10的浓度显著升高(P<0.05),Foxp3 mRNA水平明显上升(P<0.05),Treg细胞比率明显升高(P<0.05)。结论穿龙薯蓣皂苷元能明显提升DBA1/J小鼠脾脏CD4^(+)CD25^(-)T细胞上清中细胞因子IL-10浓度,Treg细胞Foxp3 mRNA水平及Treg细胞比率。

Objective To investigate the effect of diosgenin on CD4^(+)CD25^(-)T cells of DBA1/J mice.Methods CD4^(+)CD25^(-)T cells of DBA1/J mice were separated with immunomagnetic beads.Then,CD4^(+)CD25^(-)T cell purity was detected by fl ow cytometry and culture them in vitro.After treated with diosgenin,the concentration of IL-10 was detected by ELISA,the level of transcription factor Foxp3 mRNA was detected by real-time PCR,and the proportion of Treg cells was detected by FCM.Results This study separated CD4^(+)CD25^(-)T cells of 93.89%purity.Compared with the control group,the concentration of IL-10 in cell supernatant,the Foxp3 mRNA level and percentage of Treg cells were upregulated signifififi cantly(P<0.05)after treated with diosgenin for 48h.Conclusion Diosgenin can signififi cantly upregulate the levels of IL-10 and Foxp3 mRNA,the proportion of Treg cells in DBA1/J mice.