电针通过调节基质金属蛋白酶9和组织金属蛋白酶2抑制剂基因的组蛋白乙酰化,改善缺血性脑卒中后血脑屏障的破坏。

Chen Yonglin, Ouyang Ling, Meng Lingling, W U Bufan, Peng Rou, Liu Sitong, Hou Dan, Wang Yaling, Jing Xinyue, L U Shengfeng, F U Shuping
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Neurological function scores and Evans blue extravasation were employed to evaluate the poststroke injury. The effect of EA on MMP-9/TIMPs gene expression was assessed by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and chromatin immunoprecipitation (ChIP).</p><p><strong>Results: </strong>Our results showed that EA treatment prominently improved neurological function and ameliorated BBB disruption. The RT-qPCR assay showed that EA reduced the expression of MMP-9 and promoted TIMP-2 mRNA expression, but HATi reversed these effects of EA. 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引用次数: 0

摘要

目的探讨通过组蛋白乙酰化调控基质金属蛋白酶9(MMP-9)/MMPs组织抑制剂(TIMPs)基因表达是否是电针(EA)在大脑中动脉闭塞(MCAO)大鼠模型中保护血脑屏障(BBB)完整性的可能机制:雄性 Sprague-Dawley 大鼠分为四组:假组、MCAO 组、MCAO + EA(MEA)组和 MCAO + EA + HAT 抑制剂(HATi)组。MCAO 模型通过阻断大脑中动脉产生。EA应用于百汇(GV20)。再灌注后 1 或 3 d 采集样本。采用神经功能评分和埃文斯蓝外渗来评估脑卒中后的损伤。通过实时荧光定量聚合酶链反应(RT-qPCR)和染色质免疫沉淀(ChIP)评估了EA对MMP-9/TIMPs基因表达的影响:结果:我们的研究结果表明,EA治疗显著改善了神经功能,并改善了BBB的破坏。RT-qPCR测定显示,EA降低了MMP-9的表达,促进了TIMP-2 mRNA的表达,但HATi逆转了EA的这些影响。此外,ChIP结果显示,EA降低了MMP-9启动子上H3K9ace/H3K27ace的富集,并显著刺激了TIMP-2启动子上H3K9ace/H3K27ace的招募:结论:百会(GV20)EA通过组蛋白乙酰化修饰调节脑卒中急性期MMP-9和TIMP-2的转录,从而保护MCAO大鼠BBB结构的完整性。这些研究结果表明,组蛋白乙酰化介导的靶基因转录活性可能是EA治疗中风的重要机制。
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Electroacupuncture ameliorates blood-brain barrier disruption after ischemic stroke through histone acetylation regulation at the matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 2 genes.

Objective: To explore whether the regulation of matrix metalloproteinase 9 (MMP-9)/ tissue inhibitors of MMPs (TIMPs) gene expression through histone acetylation is a possible mechanism by which electroacupuncture (EA) protects blood-brain barrier (BBB) integrity in a middle cerebral artery occlusion (MCAO) rat model.

Methods: Male Sprague-Dawley rats were divided into four groups: the sham group, the MCAO group, the MCAO + EA (MEA) group, and the MCAO + EA + HAT inhibitor (HATi) group. The MCAO model was generated by blocking the middle cerebral artery. EA was applied to Baihui (GV20). Samples were collected 1 or 3 d after reperfusion. Neurological function scores and Evans blue extravasation were employed to evaluate the poststroke injury. The effect of EA on MMP-9/TIMPs gene expression was assessed by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and chromatin immunoprecipitation (ChIP).

Results: Our results showed that EA treatment prominently improved neurological function and ameliorated BBB disruption. The RT-qPCR assay showed that EA reduced the expression of MMP-9 and promoted TIMP-2 mRNA expression, but HATi reversed these effects of EA. In addition, ChIP results revealed that EA decreased the enrichment of H3K9ace/H3K27ace at MMP-9 promoters and notably stimulated the recruitment of H3K9ace/H3K27ace at TIMP-2 promoter.

Conclusion: EA treatment at Baihui (GV20) regulates the transcription of MMP-9 and TIMP-2 through histone acetylation modification in the acute stage of stroke, which preserves the structural integrity of the BBB in MCAO rats. These findings suggested that the histone acetylation-mediated transcriptional activity of target genes may be a crucial mechanism of EA treatment in stroke.

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