下调LSD1表达对于外周血Th1/Th2分化格局的影响

Effect of LSD1 on polarizing development of Th1/ Th2 in peripheral blood

目的探讨shLSD1转染人外周血CD4+T细胞后,LSD1的脱甲基酶活性对辅助性T细胞亚群1和亚群2(Th1/Th2)分化格局的影响。方法收集并利用磁珠分离纯化人外周血CD4+T细胞,PHA刺激活化48 h后,sh LSD1和化学抑制剂TCP抑制2种方法抑制LSD1的表达,采用流式细胞术和RT-PCR对人外周血T淋巴细胞亚群及CD4+T细胞功能亚型Th1、Th2的代表细胞因子IFN-γ、IL-4进行检测。结果体外分离培养外周血CD4+T细胞并用转染sh LSD1或TCP处理48 h后,流式细胞对胞内基因表达检测显示,sh LSD1和TCP组细胞IFN-γ+T细胞比例(26.13±1.89)%和(27.01±1.18)%明显高于对照组(14.67±0.65)%(P<0.05),而IL-4+T细胞比例与对照组无显著差异(P>0.05);RT-PCR检测显示,sh LSD1和TCP组IFN-γm RNA表达明显高于对照组(P<0.05),IL-4基因表达则没有改变(P>0.05)。结论LSD1可以引起CD4+T细胞向Th1/Th2分化方向改变,促进Th1方向分化,对Th2方向无明显影响。

Objective To explore the effect of LSD1 on Th1/Th2 polarizing development. Methods The peripheral blood mononuclear cells（PBMCs） from healthy donors were isolated and purified by magnetic separation anti-CD4 MACS microbeads. Stimulated by PHA for 48 hours, the expressions of IL-4 and IFN-γin peripheral blood（PBL） CD4＋T cells were analyzed by RT-PCR and fluorescence cytometry respectively. Results After TCP treatment or sh RNA-mediated knockdown of LSD1, the ratio of IFN-γ＋/IL-4＋ cells was significantly higher in sh LSD1 and TCP group than that in control group（26.13±1.89% and 27.01 ±1.18%）（P 〈0.05） by fluorescence cytometry. The IFN-γsubsets were significantly increased and the IL-4 subsets were slightly increased in sh LSD1 and TCP group than that in control group（P 〉0.05）; The expressions of IFN-γm RNA in sh LSD1 and TCP group were significantly higher than that in control group（P 〈0.05）, but the expressions of IL-4mRNA had no significant difference between the three groups（P 〉0.05）. Conclusion These findings identified that variation of Th1/Th2 cell balance of peripheral blood is correlated with LSD1 expression. Th1 cells get advantages over Th2 cells when CD4＋T cells are regulated by sh LSD1 or exposed in TCP（30 μM）.