Inhibitory effects of Baohuoside Ⅰ on the expression of FoxP3 in regulatory T cells and its mechanism of action
10.13699/j.cnki.1001-6821.2024.24.014
- VernacularTitle:宝藿苷Ⅰ抑制调节性T细胞FoxP3表达及其作用机制
- Author:
Zhao-meng CAI
1
;
Zao-yuan KUANG
1
Author Information
1. 广州中医药大学基础医学院,广东 广州 510006
- Publication Type:Journal Article
- Keywords:
Baohuoside Ⅰ;
forkhead box protein 3;
regulatory T cell;
MT-2 cell
- From:
The Chinese Journal of Clinical Pharmacology
2024;40(24):3590-3594
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects and mechanism of Baohuoside Ⅰ on transcription factors forkhead box protein 3(FoxP3)in regulatory T cell(Treg).Methods Treat induced regulatory T cells(iTreg cells:0,2,5,10,15,20 μmol·L-1 Baohuoside Ⅰ;MT-2 cells:0,2,5,10,20 μmol·L-1 Baohuoside Ⅰ)with different concentrations of Baohuoside Ⅰ for 16 hours.Use flow cytometry to detect the expression level of FoxP3,and use Western Blot to detect the expression level of FoxP3 protein.MT-2 cells were further divided into a control group and an experimental group.The control group was treated with dimethyl sulfoxide(DMSO)and 10 μg·mL-1 cycloheximide(CHX)for 0,2,4 and 6 hours;the experimental group was treated with 20 μmol·L-1Baohuoside Ⅰ and 10 μ g·mL-1 CHX for 0,2,4 and 6 hours.The degradation rate of FoxP3 in MT-2 cells from the control and experimental groups was detected by flow cytometry at different time points.Results In iTregs cells,the expression levels of FoxP3 in 0,2,5,10,15 and 20 μmol·L-1 Baohuoside Ⅰ groups were 1.00±0.00,0.98±0.07,0.95±0.04,0.86±0.04,0.57±0.04 and 0.55±0.04;the relative expression levels of FoxP3 protein were 1.00±0.00,1.04±0.14,0.86±0.06,0.64±0.33,0.36±0.15 and 0.10±0.04.The differences of above indexes between the 10,15 and 20 μmol·L-1 Baohuoside Ⅰ groups and the 0 μmol·L-1 Baohuoside Ⅰ group were statistically significant(all P＜0.001).In MT-2s cells,the expression levels of FoxP3 in0,2,5,10 and 20 μmol·L-1 Baohuoside Ⅰ groups were 1.00±0.00,0.88±0.05,0.84±0.08,0.70±0.16 and 0.57±0.14;the relative expression levels of FoxP3 protein were 1.00±0.00,0.89±0.10,0.76±0.19,0.70±0.19 and 0.37±0.12.The differences of above indexes between the 5,10 and 20 μmol·L-1 Baohuoside Ⅰ groups and the 0 μmol·L-1 Baohuoside Ⅰ group were statistically significant(P＜0.01,P＜0.001).Furthermore,Baohuoside Ⅰ decreased the stability of FoxP3 which can be observed in the CHX pulse-chase assay at the 6 hour time point where the expression levels of FoxP3 in the control group at 0.83±0.06 compared to the experimental group at 0.55±0.11(P＜0.001).Conclusion Baohuoside Ⅰcan destabilize FoxP3 in Tregs which has been shown to reduce the immunosuppressive functions of Tregs.
