Zhao Juan-Juan, Yang Shi-Wei, Zhang Fang, Yuan Xiao-Li, Zhu Zun-Min, Fang Bai-Jun, Sony Yong-Ping
Department of Hematology, Cancer Hospital Affiliated to Zhengzhou University, Henan Provincical Cancer Hospital, Zhengzhou 450008, Henan Province, China.
Department of Hematology, Peoples' Hospital of Zhengzhou University, Henan Provincial People's Hospital, Zhengzhou 450008, Henan Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2018 Jun;26(3):854-858. doi: 10.7534/j.issn.1009-2137.2018.03.037.
To investigate the effect of ATO on the proportion of Treg in the peripheral blood of patients with severe aplastic anemia (SAA) in vitro.
The peripheral blood of 20 newlydiagnosed patients were collected, and the peripheral blood monomuclear cells (PBMNC) were extracted. After the PBMNC were treated with ATO of different concentrotions (0, 1, 2.5 and 5 µmol/L) for 96 hours, the proportion of CD44 CD25CD127 regulatatory T cells (Treg) were detected by flow cytometry. The expression levels of Foxp3 mRNA were detected by RT-PCR, and the levels of IFN-γ,IL-4,IL-17 and TGF-β1 were detected by ELTSA to verify the results of flow cytomery.
ATO significantly increased the proportion of Treg (P<0.01) at the concentration of 2.5 and 5 µmol/L, and the rising degree of Treg proportion improved with the increasing ATO concentration(r= 0.524). Treg proportion increased at a concentration of 1 µmol/L, but without statistical significance (P>0.05). At 1(P<0.05), 2.5(P<0.01) and 5 µmol/L(P<0.01), ATO significantly up-regulated the expression of Foxp3 mRNA, and the increase of Foxp3 mRNA positively and linearly correlated with the increase of Treg cell-frequency(r=0.523). ATO significantly reduced the levels of IFN-γ (at ATO 1,2.5 and 5 µmol/L, P<0.01), IL-4 (at ATO 2.5 µmol/L, P<0.01; at ATO 5 µmol/L, P<0.01) and IL-17(at ATO 2.5 µmol/L, P<0.05; at ATO 5 µmol/L, P<0.01). ATO had no significant effect on TGF-β1 at 1(P>0.05) and 2.5 µmol/L (P>0.05), but significantly reduced TGF-β1 level at 5 µmol/L (P<0.05).
ATO can mediate the immune regulation through up-regulating the proportion of Treg in peripheral blood of patients with SAA and reducing the levels of IFN-γ, IL-4 and IL-17.
体外研究三氧化二砷(ATO)对重型再生障碍性贫血(SAA)患者外周血中调节性T细胞(Treg)比例的影响。
采集20例新诊断患者的外周血,提取外周血单个核细胞(PBMNC)。PBMNC用不同浓度(0、1、2.5和5 μmol/L)的ATO处理96小时后,采用流式细胞术检测CD44 CD25CD127调节性T细胞(Treg)的比例。通过逆转录聚合酶链反应(RT-PCR)检测叉头框蛋白3(Foxp3)mRNA的表达水平,采用酶联免疫吸附测定(ELTSA)检测干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)、白细胞介素-17(IL-17)和转化生长因子-β1(TGF-β1)的水平,以验证流式细胞术的结果。
在2.5和5 μmol/L浓度时,ATO显著增加Treg比例(P<0.01),且Treg比例的上升程度随ATO浓度增加而提高(r=0.524)。在1 μmol/L浓度时,Treg比例增加,但无统计学意义(P>0.05)。在1(P<0.05)、2.5(P<0.01)和5 μmol/L(P<0.01)浓度时,ATO显著上调Foxp3 mRNA的表达,且Foxp3 mRNA的增加与Treg细胞频率的增加呈正线性相关(r=0.523)。ATO显著降低IFN-γ水平(在ATO 1、2.5和5 μmol/L时,P<0.01)、IL-4水平(在ATO 2.5 μmol/L时,P<0.01;在ATO 5 μmol/L时,P<0.01)和IL-17水平(在ATO 2.5 μmol/L时,P<0.05;在ATO 5 μmol/L时,P<0.01)。在1(P>0.05)和2.5 μmol/L(P>0.05)浓度时,ATO对TGF-β1无显著影响,但在5 μmol/L浓度时显著降低TGF-β1水平(P<0.05)。
ATO可通过上调SAA患者外周血中Treg比例并降低IFN-γ、IL-4和IL-17水平来介导免疫调节。