组蛋白去乙酰化酶抑制剂SAHA对树突状细胞成熟的影响及其机制  

Effect of SAHA on Maturation of Dendritic Cells and Its Mechanism

作　　者：符明明[1] 白春洋[2] 陈昱函[1] 林进宗[1] 陆婧媛[1] 王昭[1] 张永利[3] 

机构地区：[1]厦门大学附属中山医院血液科,福建厦门361004 [2]河南大学淮河医院检验科,河南开封475000 [3]厦门大学附属中山医院急诊骨科,福建中医药大学,福建厦门361004

出　　处：《中国实验血液学杂志》2015年第3期821-825,共5页Journal of Experimental Hematology

基　　金：厦门市科技计划项目(3502Z20094012);福建省科技创新项目(2009-CXB-57)

摘　　要：目的:本研究探讨新型组蛋白去乙酰化酶抑制剂SAHA(Suberoylanilide hydroxamic acid)对健康人外周血单个核细胞(PBMNC)来源树突状细胞(DC)成熟的影响及其机制。方法:PBMNC取自健康志愿者的外周血,经贴壁处理后培养于含100 ng/ml rh GM-CSF、500 U/ml rh IL-4的RPMI 1640完全培养液中,用脂多糖(LPS)刺激经SAHA处理和未经处理的DC为实验组,并将不经LPS和SAHA处理的细胞设为对照组,在倒置显微镜下观察各组细胞形态学改变;流式细胞术检测各组DC表面抗原的表达情况;采用混合淋巴细胞培养法(MLC)观察各组DC对同种异体T淋巴细胞的刺激作用;应用凝胶电泳迁移变动测定法(EMSA)检测各组DC NF-κB通路的活化情况。结果:SAHA能够有效抑制LPS诱导的DC成熟,主要表现在经SAHA+LPS处理的DC具有未成熟DC的形态学特征;经SAHA+LPS处理组和对照组的DC表面抗原CD80、CD83、HLA-DR的表达量均低于单用LPS组(P<0.01);SAHA+LPS处理组和对照组的DC刺激同种异体淋巴细胞增殖的能力明显低于单用LPS刺激组(P<0.01);电泳迁移变动检测(EM SA)实验结果提示,SAHA+LPS处理的DC NF-κB活性显著下降。结论:SAHA能够有效抑制DC的成熟和对异基因T淋巴细胞的刺激作用,该作用与SAHA抑制DC NF-κB信号通路活化有关。Objective：To investigate the effect of SAHA on the maturation of human dendritic cells （DC） and to explore its underlying mechanism. Methods：Peripheral blood mononuclear cells （PBMNC） were isolated from human peripheral blood and cultured in RPMI 1640 medium with 100 ng/ml rhGM-CSF and 500 U/ml rhIL-4. In the LPS induced maturation process, dendritic cells treated with or without SAHA were used as test group, and dendritic cells treated without LPS or SAHA were used as control group. DC was observed under inverted microscope. Flow cytometer was used to detect the surface antigen molecules expressed by DC. The mixed lymphocyte culture （MLC） was used to observe the allogeneic lymphocyte stimulation. The NF-KB signaling pathway was detected by electrophoretic mobility shift assay （EMSA）. Results： The SAHA could effectively suppress the maturation of DC induced by LPS, the DC treated with SAHA ＋ LPS had intmature morphological characteristics; the expression of CD80, CD83 and HLA-DR in SAHA ＋ LPS group and control group were significantly down-regulated as compared with single LPS group （ P 〈 0. 01 ） ; the ability of DC to stimulate the proliferation of allogeneic T lymphocytes in SAHA ＋ LPS group and control group was significantly weaker than that in single LPS group （ P 〈 0.01 ） ; EMSA results showed that NF-KB activity decreased after SAHA and LPS treatment and was significantly lower than that of single LPS group. Conclusion： SAHA can effectively suppress DC maturation induced by LPS and also weaken the ability to stimulate allogeneic T lymphocyte. NF-KB signaling pathway is involved in regulating DC maturation.

关 键 词：组蛋白去乙酰化酶抑制剂 SAHA 树突状细胞 NF-ΚB 

分 类 号：R392.1[医药卫生—免疫学]