Epigenetic Activation of Circadian Clock Genes Elicits Inflammation in Experimental Murine Dry Eye.
Epigenetic Activation of Circadian Clock Genes Elicits Inflammation in Experimental Murine Dry Eye.
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DOI：
10.1080/09273948.2023.2205525
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发表时间：
2023-05
影响因子：
3.3
通讯作者：
Ruifen Wei;Qianqian Chen;Qinxiang Zheng;P. Reinach;Xiying Tan;Chengjie Pan;Wei Xu;L. Tong
中科院分区：
文献类型：
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作者：
Ruifen Wei;Qianqian Chen;Qinxiang Zheng;P. Reinach;Xiying Tan;Chengjie Pan;Wei Xu;L. Tong
目的探讨生物钟基因在实验性干眼(EDE)炎症反应中的作用。方法采用RNA测序的方法，分析EDE模型中的mRNA表达模式。RT-qPCR和/或Western印迹检测EDE过程中炎症因子和昼夜节律基因的表达。甲基靶™检测体内PER基因启动子甲基化水平。PER2或PER3基因敲除在体外评估了它们对炎性因子的影响。结果利用智能控制环境系统(ICES)建立了小鼠EDE模型。显著上调的基因丰富了昼夜节律。在EDE过程中，PER2和PER3及其启动子的低甲基化存在振荡和时间依赖性的上调。在体外模型中，沉默PER2或PER3显著降低炎症因子的表达，并逆转这种增加的炎症反应。结论DNA甲基化介导了PER2和PER3的上调，导致了EDE的炎症反应。
PURPOSE To explore whether circadian clock genes contribute to elicit inflammation in experimental dry eye (EDE). METHODS RNA sequencing analyzed mRNA expression patterns in EDE model. RT-qPCR and/or Western blot determined the expression of inflammatory factors and circadian genes during EDE. MethylTarget™ assays determined the promoter methylation levels of Per genes in vivo. Per2 or Per3 knockdown assessed their effects on inflammatory factors in vitro. RESULTS We utilized an intelligently controlled environmental system (ICES) to establish a mouse EDE model. The significant upregulated genes were enriched for circadian rhythms. Therein lied oscillatory and time-dependent upregulation of PER2 and PER3, as well as their promoter hypomethylation during EDE. Silencing PER2 or PER3 significantly decreased inflammatory factor expression and also reversed such increased inflammatory response in azacitidine (AZA) treatment in vitro model. CONCLUSIONS Our findings suggest that DNA methylation mediated the upregulation of PER2 and PER3, leading to inflammatory response in EDE.