Endothelial Cells retain inflammatory memory through chromatin remodeling [ATAC-seq]

Sepsis survivors exhibit long-term endothelial dysfunction, increasing susceptibility to secondary infections such as pneumonia. To investigate the epigenetic and transcriptional basis of endothelial inflammatory memory, we employed transcriptomic profiling of primary endothelial cells exposed to inflammatory stimuli in a two-hit model. This study reveals persistent transcriptional reprogramming following IL-6 exposure and identifies a primed endothelial state that amplifies responses to secondary challenges. Overall design: Primary endothelial cells were treated with IL-6 or PBS for 72 hours (first hit), followed by a 48-hour washout phase. Nuclei were isolated for ATAC-seq from cells collected at two timepoints: immediately after the 72-hour treatment (IL6_72h, PBS_72h) and after the 48-hour washout (IL6_72h_W48h, PBS_72h_W48h). Each group included two or three biological replicates

脓毒症幸存者会出现长期内皮功能障碍（endothelial dysfunction），从而增加罹患肺炎等继发性感染的易感性。为探究内皮炎症记忆的表观遗传（epigenetic）与转录调控基础，我们采用双打击模型（two-hit model），对经炎症刺激的原代内皮细胞（primary endothelial cells）开展转录组谱分析（transcriptomic profiling）。本研究揭示了白细胞介素6（IL-6）刺激后持续存在的转录重编程（transcriptional reprogramming）现象，并鉴定出一种可放大继发性刺激应答的致敏内皮状态。总体实验设计：将原代内皮细胞用白细胞介素6（IL-6）或磷酸盐缓冲液（PBS）处理72小时（首次打击），随后进行48小时的洗脱培养。分别在两个时间点收集细胞并分离细胞核，开展转座酶可及性染色质测序（ATAC-seq）分析：一是72小时处理结束即刻（IL6_72h、PBS_72h），二是48小时洗脱培养结束后（IL6_72h_W48h、PBS_72h_W48h）。每组设置2至3个生物学重复。