Department of Surgical Gastroenterology, Rigshospitalet, University of Copenhagen, Denmark; Center for Surgical Translational and Artificial Intelligence Research (CSTAR), Rigshospitalet, University of Copenhagen, Denmark.
Department of Anesthesia, Center of Head and Orthopedics, Rigshospitalet, University of Copenhagen, Denmark.
Injury. 2021 Aug;52(8):2095-2103. doi: 10.1016/j.injury.2021.03.050. Epub 2021 Mar 26.
Epigenetic changes have been described in trauma patients in the form of histone acetylation events, but whether DNA-methylation occurs remains unknown. We hypothesized that the combination of hemorrhage and saline resuscitation would alter DNA-methylation and associated proteomic profiles in the rat lung.
Ten rats were subjected to a pressure-controlled hemorrhage and resuscitation model consisting of hemorrhage to a mean arterial pressure (MAP) of 35mmHg for 90 minutes, followed by saline resuscitation to a MAP >70mmHg for 90 minutes (n=5) or sham (only anesthesia and cannulation). Lungs were harvested and subjected to reduced genome wide DNA-methylation analysis through bisulphite sequencing as well as proteomics analysis. Data was analyzed for differentially methylated regions and associated alterations in proteomic networks through a weighted correlation network analysis (WCNA). Pathway analysis was used to establish biological relevance of findings.
Hemorrhage and saline resuscitation were associated with differential methylation of 353 sites across the genome compared to the sham group. Of these, 30 were localized to gene promoter regions, 31 to exon regions and 87 to intron regions. Network analysis identified an association between hemorrhage/resuscitation and DNA-methylation events located to genes involved in areas of endothelial and immune response signaling. The associated proteomic response was characterized by activations of mRNA processing as well as endothelial Nitric Oxide Synthase (eNOS) metabolism.
We demonstrated an association between DNA-methylation and hemorrhage/saline resuscitation. These results suggest a potential role of DNA-methylation in the host response to injury.
创伤患者的组蛋白乙酰化事件表现出表观遗传改变，但 DNA 甲基化是否发生尚不清楚。我们假设出血和盐水复苏的组合会改变大鼠肺中的 DNA 甲基化和相关蛋白质组学特征。
10 只大鼠接受了一种压力控制的出血和复苏模型，包括出血至平均动脉压 (MAP) 为 35mmHg 持续 90 分钟，然后用盐水复苏至 MAP > 70mmHg 持续 90 分钟（n=5）或假手术（仅麻醉和插管）。收获肺部，并通过亚硫酸氢盐测序进行全基因组范围的 DNA 甲基化分析，以及蛋白质组学分析。通过加权相关网络分析 (WCNA) 分析差异甲基化区域和相关蛋白质组网络的变化。通路分析用于确定发现的生物学相关性。
与假手术组相比，出血和盐水复苏与全基因组 353 个位点的差异甲基化相关。其中，30 个位于基因启动子区域，31 个位于外显子区域，87 个位于内含子区域。网络分析确定了出血/复苏与位于内皮和免疫反应信号转导相关基因中的 DNA 甲基化事件之间的关联。相关的蛋白质组反应特征是 mRNA 处理以及内皮型一氧化氮合酶 (eNOS) 代谢的激活。
我们证明了 DNA 甲基化与出血/盐水复苏之间的关联。这些结果表明 DNA 甲基化在宿主对损伤的反应中可能发挥作用。
