Acute liver failure (ALF) is a severe life-threatening disease associated with the disorder of the gut-liver axis. However, the cellular characteristics of ALF in the gut and related therapeutic targets remain unexplored. Here, we utilized the D-GALN/LPS (D/L)-induced ALF model to characterize 33,216 single-cell transcriptomes and define a mouse ALF intestinal cellular atlas. We found that unique, previously uncharacterized intestinal immune cells, including T cells, B cells, macrophages, and neutrophils, are responsive to ALF, and we identified the transcriptional profiles of these subsets during ALF. We also delineated the heterogeneity of intestinal epithelial cells (IECs) and found that ALF-induced cell cycle arrest in intestinal stem cells and activated specific enterocyte and goblet cell clusters. Notably, the most significantly altered IECs, including enterocytes, intestinal stem cells and goblet cells, had similar activation patterns closely associated with inflammation from intestinal immune activation. Furthermore, our results unveiled a common Ep300-dependent transcriptional program that coordinates IEC activation during ALF, which was confirmed to be universal in different ALF models. Pharmacological inhibition of Ep300 with an inhibitor (SGC-CBP30) inhibited this cell-specific program, confirming that Ep300 is an effective target for alleviating ALF. Mechanistically, Ep300 inhibition restrained inflammation and oxidative stress in the dysregulated cluster of IECs through the P38-JNK pathway and corrected intestinal ecology by regulating intestinal microbial composition and metabolism, thereby protecting IECs and attenuating ALF. These findings confirm that Ep300 is a novel therapeutic target in ALF and pave the way for future pathophysiological studies on ALF.

急性肝衰竭（ALF）是一种与肠肝轴紊乱相关的严重危及生命的疾病。然而，肠道 ALF 的细胞特征和相关治疗靶点仍有待探索。在这里，我们利用 D-GALN/LPS (D/L) 诱导的 ALF 模型来表征 33,216 个单细胞转录组，并定义小鼠 ALF 肠道细胞图谱。我们发现独特的、以前未表征的肠道免疫细胞，包括 T 细胞、B 细胞、巨噬细胞和中性粒细胞，对 ALF 有反应，并且我们确定了 ALF 期间这些亚群的转录谱。我们还描述了肠上皮细胞 (IEC) 的异质性，发现 ALF 诱导肠干细胞细胞周期停滞，并激活特定的肠上皮细胞和杯状细胞簇。值得注意的是，改变最显着的 IEC，包括肠上皮细胞、肠干细胞和杯状细胞，具有与肠道免疫激活引起的炎症密切相关的相似激活模式。此外，我们的结果揭示了一个常见的Ep300依赖性转录程序，该程序在 ALF 期间协调 IEC 激活，该程序被证实在不同的 ALF 模型中具有通用性。用抑制剂（SGC-CBP30）对Ep300进行药理学抑制可抑制该细胞特异性程序，证实Ep300是缓解 ALF 的有效靶点。从机制上讲，Ep300抑制通过P38-JNK通路抑制失调的IEC簇中的炎症和氧化应激，并通过调节肠道微生物组成和代谢来纠正肠道生态，从而保护IEC并减弱ALF。这些发现证实Ep300是ALF的一个新的治疗靶点，并为未来ALF的病理生理学研究铺平了道路。