摘要目的 探讨支气管哮喘(简称哮喘)患者外周血组蛋白去乙酰化酶9 (Histone Deacetylase 9,HDAC9)的表达特征及其与Th2、Th17、调节性T细胞的关系.方法 选择2010年3月至2011年6月上海瑞金医院呼吸科47例慢性持续期哮喘患者,分为间歇期哮喘组、轻度哮喘组和中重度哮喘组,各组均行肺功能检查和哮喘控制问卷调查哮喘控制情况.并选取20名健康人作为健康对照组.通过分离外周血单个核细胞(PBMCs),Trizol法抽提RNA,采用荧光实时定量方法检测转录因子GATA3、IL-4、HDAC9mRNA表达水平.ELISA检测血浆中Th17细胞分泌的主要细胞因子IL-17以及调节性T细胞分泌的主要细胞因子——转化生长因子( TGF)-β表达水平.结果 哮喘组PBMCs中GATA3、IL-4mRNA总体相对表达水平分别为28.12±7.57、743.6±312.8高于健康对照组相对表达水平(分别为0.56±0.22、0.7±0.8),差异有统计学意义(U值分别为16.00和37.00,P ＜0.01),HDAC9 mRNA在间歇期、轻度、中重度哮喘患者中的表达水平分别为3.20±0.50、89.6±18.0和323.0±65.3,呈逐渐上升趋势,各组间差异有统计学意义(H=11.32,P＜0.05).同时,哮喘患者总体血浆细胞因子IL-17表达水平为83±55,高于健康对照组的34±22(U=153.50,P＜0.01),而TGF-β表达水平下降趋势,组间无统计学差异.HDAC9 mRNA表达水平与GATA3 mRNA表达水平呈正相关(r =0.482,P＜0.05),与IL-4 mRNA表达水平呈正相关(r=0.432,P＜0.05),与IL-17水平呈现正相关(r=0.538,P＜0.05),与TGF-β表达水平呈负相关(r=-0.417,P＜0.05).在中重度哮喘患者中,HDAC9mRNA水平与肺功能FEV1占预计值％呈负相关(r=-0.657,P＜0.05).结论 哮喘患者PBMCs中HDAC9mRNA表达水平随病情严重程度逐渐升高,其升高不仅与哮喘发病相关的Th2细胞特征性转录因子GATA3、细胞因子IL-4 mRNA表达水平以及Th17、Treg分泌的细胞因子水平密切相关,且与哮喘患者FEV1占预计值％密切相关.

abstractsObjective To investigate the expression pattern of histone deacetylase 9 (HDAC9) in peripheral blood of asthmatics and its effect on immune cells (Th2,Th17,Tregs ) involved in the pathogenesis of asthma.Methods Forty-seven asthmatics from Ruijin Hospital were recruited and assigned to intermittent,mild and moderate-severe groups.Lung function test and Asthma Control Questionnaire were performed to evaluate asthma control and severity.Twenty healthy donors were enrolled as controls.GATA3,IL-4,and HDAC9 mRNA expression levels were measured by SYRB Green Real-time PCR.The cytokine IL-17-mainly produced by Th17 cells and TGF-β-mainly produced by Treg cells,were measured by ELISA.Results The GATA3 and IL-4 mRNA expression levels ( 28.12 ± 7.57 and 743.6 ± 312.8 ) were upregulated in asthmatics as compared to the healthy controls [ 0.56 ± 0.22,0.7 ± 0.8 ( U =16.00,37.00,P＜0.01 )].The HDAC9 mRNA expression levels of intermittent,mild and moderate-severe groups were 3.20 ± 0.50,89.6 ± 18.0,323.0 ± 65.3,respectively,which were associated with the severity of disease (H =11.32,P ＜0.05).The level of IL-17 in asthmatic group was(83 ±55)ng/L,which was up-regulated as compared to the healthy control [ (34 ± 22 ) ng/L ( U =153.50,P ＜ 0.01 ) ].The level of TGF-β was decreased in the asthmatic groups as compared to the healthy control,but the difference did not reach significance.HDAC9 mRNA expression level was positively correlated with GATA3 mRNA expression level (r =0.482,P ＜0.05),and also with IL-4 mRNA expression (r =0.432,P＜0.05) and IL-17 (r=0.538,P ＜ 0.05 ),but negatively correlated with TGF-β ( r =- 0.417,P ＜ 0.05 ).In patients with moderate-severe asthma,HDAC9 mRNA expression level was negatively correlated with FEV1％ (r =-0.657,P ＜ 0.05 ).Conclusion HDAC9 mRNA expression was up-regulated in peripheral blood of asthmatics,which was not only associate with the Th2 master transcriptional factors GATA3,cytokine IL-4 mRNA,Thl7 and Treg cell-related cytokines,but also with FEV1％ in moderate-severe asthma.