Hou Xiaoxia, Wan Huanying, Ai Xiangyan, Shi Yuheng, Ni Yingmeng, Tang Wei, Shi Guochao
Department of Pulmonary Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Clin Respir J. 2016 May;10(3):371-9. doi: 10.1111/crj.12227. Epub 2014 Nov 9.
The aim of this study is to investigate the expression pattern of histone deacetylase 9 in peripheral blood of patients with allergic asthma and its regulatory effect on the balance of Th17/Treg cells involved in the pathogenesis of asthma.
flap-Ub promoter-GFP-WRE vector was used to construct the Jurkat-HA-FOXP3 cell line. After histone deacetylase inhibitor-trichostatin A (TSA) treatment, FOXP3 and RORγt expression were detected by real-time-polymerase chain reaction (RT-PCR). BALB/c mice were randomly assigned to control group, TSA treatment and the asthma group. Serum Immunoglobulin E (IgE) was detected with enzyme-linked immunosorbent assay (ELISA), airway inflammation in lung tissue evaluated by haematoxylin/eosin staining, bronchoalveolar lavage fluid (BALF) cell number and differential counted, interleukin (IL)-17A and TGF-β concentrations in BALF measured with ELISA, and expression of RORγt and FOXP3 messenger RNA (mRNA)measured by RT-PCR. Forty-seven patients with asthma were recruited and assigned to intermittent, mild and moderate-severe group. GATA3, IL-4, histone deacetylases (HDAC) 9 mRNA expression level were measured by RT-PCR.
After TSA treatment, FOXP3 mRNA level was upregulated, while RORγt mRNA level was downregulated. FOXP3 protein level was also upregulated by TSA. In vivo, TSA treatment can inhibit IL-17 but promote transforming growth factor-beta production in the BALF of asthma mice, and inhibited the expression of Th17 cells and RORγt mRNA in lung; also can promote Foxp3 mRNA expression. GATA3, IL-4 mRNA expression levels were upregulated in patients with asthma than the healthy control. HDAC9 mRNA expression level was associated with the severity of disease.
The histone deacetylase inhibitor TSA can regulate the balance of Th17/Treg in asthma by regulating the activity of histone deacetylase.
本研究旨在探讨组蛋白去乙酰化酶9在过敏性哮喘患者外周血中的表达模式及其对哮喘发病机制中Th17/Treg细胞平衡的调节作用。
采用flap-Ub启动子-GFP-WRE载体构建Jurkat-HA-FOXP3细胞系。经组蛋白去乙酰化酶抑制剂曲古抑菌素A（TSA）处理后，通过实时聚合酶链反应（RT-PCR）检测FOXP3和RORγt的表达。将BALB/c小鼠随机分为对照组、TSA处理组和哮喘组。采用酶联免疫吸附测定（ELISA）检测血清免疫球蛋白E（IgE），通过苏木精/伊红染色评估肺组织气道炎症，对支气管肺泡灌洗液（BALF）细胞进行计数和分类，用ELISA检测BALF中白细胞介素（IL）-17A和转化生长因子-β（TGF-β）的浓度，用RT-PCR检测RORγt和FOXP3信使核糖核酸（mRNA）的表达。招募47例哮喘患者并分为间歇组、轻度组和中度-重度组。采用RT-PCR检测GATA3、IL-4、组蛋白去乙酰化酶（HDAC）9 mRNA表达水平。
TSA处理后，FOXP3 mRNA水平上调，而RORγt mRNA水平下调。TSA还上调了FOXP3蛋白水平。在体内，TSA处理可抑制哮喘小鼠BALF中IL-17的产生，但促进TGF-β的产生，并抑制肺中Th17细胞和RORγt mRNA的表达；还可促进Foxp3 mRNA表达。哮喘患者GATA3、IL-4 mRNA表达水平高于健康对照。HDAC9 mRNA表达水平与疾病严重程度相关。
组蛋白去乙酰化酶抑制剂TSA可通过调节组蛋白去乙酰化酶的活性来调节哮喘中Th17/Treg的平衡。
