组蛋白去乙酰化酶抑制剂Belinostat对髓源性树突状细胞免疫功能的作用研究  

Study on the immune functions of dendritic cells regulated by histone deacetylase inhibitor Belinostat

作　　者：贾文华 毛慧 陈婉如 岳晓彤 魏鑫鑫 李德鹏 徐开林 黄一虹 

机构地区：[1]徐州医科大学附属医院血液科,221002 [2]西安医学院第一附属医院血液科,710077 [3]徐州医科大学附属第三医院血液科,221003 [4]徐州医科大学血液病研究所

出　　处：《中华血液学杂志》2018年第1期41-46,共6页Chinese Journal of Hematology

基　　金：江苏省卫生科研项目（H201427）;徐州市科技计划（KC16SH016）;江苏省2015.2016年度普通高校研究生科研创新计划（SJLX15_0722、SJZZ16_0288）

摘　　要：目的探讨组蛋白去乙酰化酶（HDAC）抑制剂Belinostat对小鼠骨髓来源树突状细胞（DC）免疫功能的影响及初步机制。方法体外诱导培养C57BL／6小鼠骨髓来源的DC，诱导培养第5天为未成熟DC（imDC）组，设0、50、100nmol／LBelinostat作用组；imDC以脂多糖作用24h为成熟DC（mDC），设0、50、100nmol／LBelinostat作用组。从细胞形态、超微结构、免疫表型进行鉴定。流式细胞术检测各组DC免疫表型及趋化因子受体CCR7表达水平，趋化实验检测DC的体外迁移率。单向混合淋巴细胞培养法检测各组DC刺激下异基因淋巴细胞增殖率。ELISA法检测各组DC培养上清中TNF—α、IL-12及IL-10的表达水平。RQ—PCR检测Belinostat对DC中RelBmRNA表达水平的影响。结果成功诱导培养出imDC及mDC并鉴定。50和100nmol／LBelinostat＋imDC组CCR7表达水平均低于imDc组[（25．82±7．25）％对（50．44±5．61）％、（18．71±2．00）％对（50．44±5．61）％j；50nmol／LBelinostat＋mDC组CCR7表达水平高于mDC组[（71．14±1．96）％对（64．90±1．47）％]。Belinostat作用下imDC和mDC的迁移率均下降，但在imDC中组间比较差异无统计学意义。当刺激细胞：反应细胞比例为1：2时，100nmol／LBelinostat＋imDC刺激下淋巴细胞增殖率低于imDC组[（227．09±13．49）％对（309．49±53．69）％]。Belinostat作用下mDC所分泌的TNF-α、IL-12和IL-10较mDC组均明显下降，差异均有统计学意义（P值均〈0．01）。Belinostat＋imDC及Belinostat＋mDC中RelBmRNA表达水平较imDC组及mDC组均有降低（P值均〈0．05）。结论Belinostat可调节DC迁移、抑制T淋巴细胞增殖及细胞因子分泌，一定程度上抑制DC成熟，可能与其下调DC中NF—kB的转录因子RelBmRNA水平有关。Objective To explore effects of histone deacetylase inhibitor Belinostat on the immunologic function of dendritic cells （DC） and its possible mechanism. Methods Cultured mouse bone marrow-derived DC from C57BL/6 mouse in vitro. The experiments were divided into 0, 50, 100 nmol/L Belinostat ＋ immature DC （imDC） group, and 0, 50, 100 nmol/L Belinostat mature DC （mDC）. The changes of the ultrastructure of DC were observed by transmission electron microscope （TEM）. Immunophenotype and CCR7 expression rate were detected by FCM, and the migration rate was observed by chemotaxis assay. The proliferation of lymphocytes stimulated by different DC was detected by mixed lymphocyte culture reaction. The cytokines in the culture supernatant, including TNF-ct, IL-12 and IL-10, were examined by ELISA. RQ-PCR was used to examine the relative expression of mRNA in RelB. Results Successful cultured and identified the qualified imDC and mDC. Belinostat decreased the expression of CCR7 on imDC [（25.82±7.25）% vs （50.44±5.61）% and 08.71±2.00）% vs （50.44±5.61）%],meanwhile increased the rate on mDC [（71.14±1.96）% vs （64.90± 1.47）%]. Chemotaxis assay showed that the migration rate of Belinostat＋imDC and Belinostat＋mDC group were both decreased, but the difference in imDC was not significant. T lymphocyte proliferation rate stimulated by 100 nmol/L Belinostat＋imDC group was lower than imDC group in condition irritation cell： reaction cell= 1：2 [（227.09± 13.49）% vs （309.49±53.69）%]. Belinostat significantly suppressed the secretion of cytokines TNF-α, IL-12 and IL-10 （all P 〈 0.01）. The relative expression of mRNA in RelB was slightly decreased in Belinostat＋imDC and Belinostat＋mDC group （all P 〈 0.05）. Conclusion Belinostat could effectly suppress DC maturation and regulate immune tolerance of DC, which may be due to the down-regulation of mRNA level of RelB in DC.

关 键 词：组蛋白去乙酰化酶抑制剂 Belinostat 树突细胞 免疫耐受 

分 类 号：R392[医药卫生—免疫学]