组蛋白乙酰转移酶MOF在多发性硬化发病机制中的作用

doi:10.6040/j.issn.1671-7554.0.2015.366

摘要/Abstract

摘要： 目的探讨实验性自身免疫性脑脊髓炎(EAE)小鼠发病高峰期、缓解期Treg细胞和组蛋白乙酰转移酶MOF表达的变化情况,以及MOF对EAE发病机制的影响。方法将20只体质量相近的6~8周龄C57BL/6雌鼠随机分为两组,EAE组第0天腹股沟处皮下注射200 μg MOG_35-55,对照组注射等量弗氏完全佐剂,两组均腹腔注射200 ng百日咳毒素,第2天腹腔注射200 ng 百日咳毒素。取EAE组不同发病时期和对照组小鼠的脾脏组织。Western blotting法检测MOF、FOXP3和H4K16ac蛋白的表达情况;qRT-PCR检测脾脏组织中MOF、FOXP3、TLRS mRNA的表达水平;蛋白质免疫共沉淀(CoIP)法检测MOF与FOXP3在脾脏组织中有无相互作用;染色质免疫共沉淀(ChIP-qPCR)法检测MOF的下游靶基因;在HCT116细胞中过表达MOF后检测FOXP3、 TLRS表达的变化情况。结果 EAE组脾脏中的MOF及FOXP3蛋白表达水平显著高于对照组;EAE组小鼠脾脏中MOF、FOXP3、IL17、TLR4、TLR5、TLR6、TLR7、TLR9 mRNA水平显著高于对照组(P<0.05);MOF与FOXP3在脾脏组织中结合并有相互作用;脾脏组织中,MOF与FOXP3、TLR3、TLR4、TLR5、TLR6、SMAD2、SMAD3和RoRγt启动子结合;MOF过表达后,TLR4和FOXP3表达量增加(P<0.05)。结论 MOF通过与FOXP3相互作用,抑制EAE的发生。

关键词: 组蛋白乙酰转移酶, MOF, Toll样受体, FOXP3, Treg

Abstract: Objective To investigate the activities of Treg cells and the expression of MOF in the peak and remission periods of experimental allergic encephalomyelitis(EAE)mice and the impact of MOF on the pathogenesis of EAE. Methods On Day 0, twenty 6-8 week old C57BL/6 female mice of similar weight were subcutaneously injected with 200 μg MOG_35-55 or complete freunds adjuvant(CFA)and then intraperitoneally injected with 200 ng pertussis toxin. On Day 2, the above three mice were intraperitoneally injected with 200 ng pertussis toxin. The spleen tissues of the EAE mice in different phases were used, along with the spleen tissues of the mice from control group in the same phases. The protein levels of MOF, FOXP3, and H4K16ac were detected through Western blotting. The mRNA expression levels of MOF, TLRS, and FOXP3 were detected by real-time fluorescent quantitative RT-PCR. Co-immunoprecipitation was performed to assess the binding between MOF and FOXP3 in the spleen tissues. ChIP-qPCR was performed to detect the downstream target genes of MOF. The relationships between the protein expression levels of TLRS, FOXP3 in the HCT116 cells, and MOF regulation were also studied. Results The expression levels of MOF and FOXP3 protein in the spleen of EAE mice were significantly higher than those of the control group. The mRNA levels of FOXP3, 山东大学学报 (医学版)54卷2期 -关景云,等.组蛋白乙酰转移酶MOF在多发性硬化发病机制中的作用 \=-MOF, IL17, TLR4, TLR5, TLR6, TLR7 and TLR9 in EAE mouse spleen tissue were significantly higher than those of the control group(P<0.05); Co-IP showed that MOF directly interacted with FOXP3 in spleen tissues; ChIP-qPCR showed that MOF combined with FOXP3, TLR3, TLR4, TLR5, TLR6, SMAD2, SMAD3, and RoRγt genes in the spleen. TLR4 and FOXP3 showed higher expression levels in the HCT116 cells compared with the control group when MOF was overexpressed(P<0.05). Conclusion MOF suppresses the occurrence of EAE through its interaction with FOXP3.

Key words: Histone acetyltransferase, Treg, Toll-like receptors, MOF, FOXP3

中图分类号:

R392.8

关景云,杨阳,邱春红,李相芝. 组蛋白乙酰转移酶MOF在多发性硬化发病机制中的作用[J]. 山东大学学报（医学版）, 2016, 54(2): 27-32.

GUAN Jingyun, YANG Yang, QIU Chunhong, LI Xiangzhi. Role of histone acetyltransferase MOF in the pathogenesis of multiple sclerosis[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(2): 27-32.

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