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目的观察高迁移率族蛋白B1(high mobility group box-1 protein,HMGB1)对调节性T细胞(regulatory Tcell,Treg)叉头翼状螺旋转录因子(forkhead/winged helix transcription factorp3,Foxp3)基因及蛋白表达的影响,并对其机制进行初步探讨。方法免疫磁珠法分离正常BALB/c小鼠脾脏Treg。采用固相包被抗-CD3及可溶性CD28辅助活化,给予HMGB1刺激,观察HMGB1刺激与Foxp3基因及蛋白表达的时间-效应关系及剂量-效应关系。结果经HMGB1刺激后的TregFoxp3蛋白表达于24～72h明显下调(P<0.05,P<0.01),其中以作用48、72h后表达下调尤为显著(P<0.01);给予不同剂量HMGB1刺激72h后,10、100、1000ng/ml的HMGB1均可诱导Foxp3表达减弱(P<0.05,P<0.01),其中HMGB1的浓度在1000ng/ml时Foxp3表达下调最明显。Foxp3mRNA表达呈现出与蛋白表达相同的时间、剂量依赖关系。结论HMGB1通过诱导TregFoxp3mRNA表达下调,进一步影响其蛋白产物合成,从而影响Treg免疫调节活性。
Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on the gene and protein expression of forkhead / winged transcription factor 3 (Foxp3) in regulatory T cell (Treg) The impact of its mechanism and conduct a preliminary discussion. Methods Immunomagnetic beads method was used to isolate Treg from normal BALB / c mice. The solid phase coated anti-CD3 and soluble CD28 were used to assist the activation. The cells were stimulated with HMGB1 to observe the time-effect relationship and dose-response relationship between HMGB1 stimulation and Foxp3 gene and protein expression. Results The expression of TregFoxp3 protein was significantly down-regulated after HMGB1 stimulation for 24-72h (P <0.05, P <0.01), especially after 48 and 72h treatment (P <0.01). After stimulated with different doses of HMGB1 for 72h, HMGB1 at 10, 100 and 1000ng / ml could reduce the expression of Foxp3 (P <0.05, P <0.01), and the down-regulation of Foxp3 was the most significant at HMGB1 concentration of 1000ng / ml. Foxp3 mRNA expression showed the same time and protein expression in a dose-dependent manner. Conclusion HMGB1 can down-regulate the expression of Treg Foxp3 mRNA and further affect its protein synthesis, thereby affecting the immunoregulatory activity of Treg.