Silencing of microRNA-494 inhibits the neurotoxic Th1 shift via regulating HDAC2-STAT4 cascade in ischaemic stroke
Silencing of microRNA-494 inhibits the neurotoxic Th1 shift via regulating HDAC2-STAT4 cascade in ischaemic stroke
复制标题
沉默 microRNA-494 通过调节缺血性中风中的 HDAC2-STAT4 级联抑制神经毒性 Th1 转变
DOI：
10.1111/bph.14852
复制
发表时间：
2019-11-08
影响因子：
7.3
通讯作者：
Luo, Yumin
中科院分区：
文献类型：
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作者：
Zhao, Haiping;Li, Guangwen;Luo, Yumin
背景和目的T辅助细胞1(Th1)偏斜的神经毒性是导致啮齿动物卒中预后不良的原因之一。在这里，我们阐明了急性缺血性中风(AIS)患者超急性期Th1/Th2转变的机制，并寻找基于miRNA的治疗靶点。实验方法：用实时荧光定量聚合酶链式反应检测AIS患者和对照组血液中MIR-494的水平。C57BL/6J小鼠采用短暂性大脑中动脉闭塞模型，大脑皮质神经元缺氧缺糖。用荧光素酶报告系统、染色质免疫沉淀测序(CHIP-SEQ)和CHIP-PCR来揭示可能的机制。主要结果：AIS患者淋巴细胞中Th1/Th2发生移位，组蛋白脱乙酰基酶2(HDAC2)表达明显下调。CHIP-SEQ显示HDAC2结合位点富集调节Th1细胞因子的产生，CHIP-PCR证实在AIS患者淋巴细胞的STAT4内含子和T-box转录因子21(T-bet)启动子处的HDAC2结合发生了改变。MIR-494是AIS患者淋巴细胞中最显著增加的miRNA，miR-494-3p直接靶向HDAC2。在AIS患者中，miR-494和Th1细胞因子与美国国立卫生研究院卒中量表(NIHSS)测量的神经功能缺陷之间存在很强的相关性。体外和体内实验表明，antagomir-494通过HDAC2-STAT4途径减轻了Th1移位介导的缺血性卒中小鼠神经元和感觉运动功能的损伤。结论和意义我们证明，miR-494抑制通过调节HDAC2-STAT4级联来防止Th1偏斜的神经毒性。
Background and Purpose T helper cell 1 (Th1)-skewed neurotoxicity contributes to the poor outcome of stroke in rodents. Here, we have elucidated the mechanism of the Th1/Th2 shift in acute ischaemic stroke (AIS) patients at hyperacute phase and have looked for a miRNA-based therapeutic target. Experimental Approach MiR-494 levels in blood from AIS patients and controls were measured by real-time PCR. C57BL/6J mice were subjected to transient middle cerebral artery occlusion, and cortical neurons were subjected to oxygen-glucose deprivation. Luciferase reporter system, chromatin immunoprecipitation sequencing (ChIP-Seq), and ChIP-PCR were used to uncover possible mechanisms. Key Results In lymphocytes from AIS patients, there was a Th1/Th2 shift and histone deacetylase 2 (HDAC2) was markedly down-regulated. ChIP-seq showed that HDAC2 binding sites were enriched in regulation of Th1 cytokine production, and ChIP-PCR confirmed that HDAC2 binding was changed at the intron of STAT4 and the promoter of T-box transcription factor 21 (T-bet) in lymphocytes from AIS patients. MiR-494 was the most significantly increased miRNA in lymphocytes from AIS patients, and miR-494-3p directly targeted HDAC2. A strong association existed between miR-494 and Th1 cytokines, and neurological deficit as measured by the National Institute of Health Stroke Scale (NIHSS) in AIS patients. In vitro and in vivo experiments showed that antagomir-494 reduced Th1 shift-mediated neuronal and sensorimotor functional damage in the mouse model of ischaemic stroke, via the HDAC2-STAT4 pathway. Conclusion and Implications We demonstrated that miR-494 inhibition prevented Th1-skewed neurotoxicity through regulation of the HDAC2-STAT4 cascade.