背景:急性肝衰竭(ALF)是一种威胁生命的疾病,由自限性急性肝损伤(ALI)发展而来。以窦状高凝和随后的大量缺氧肝细胞损伤为特征的微循环障碍被认为是ALI恶化为ALF的机制;然而,正弦高凝的确切分子途径仍然未知.这里,我们分析了ALI患者和小鼠模型,以揭示ALI伴有微循环障碍的发病机制。
方法:我们对ALI进行了单中心回顾性研究,并分析了血液样本和肝组织以评估ALI患者(n=120)的微循环障碍。将单细胞RNA测序分析(scRNA-seq)应用于来自伴刀豆球蛋白A(ConA)诱导的ALI小鼠模型的肝脏。干扰素-γ(IFNγ)和肿瘤坏死因子-α基因敲除小鼠,和原代人肝窦内皮细胞(LSECs)用于评估微循环障碍的机制。
结果:有微循环障碍的ALI患者血清IFNγ浓度明显高于无微循环障碍的患者,在出现微循环障碍的ConA小鼠模型中,IFNγ上调。肝IFNγ表达早在ConA治疗后1小时就增加,然后才出现正弦高凝和缺氧性肝损伤。scRNA-seq显示IFNγ在先天性淋巴细胞中上调,并在肝损伤早期刺激肝血管内皮细胞。在用ConA处理的IFNγ敲除小鼠中,肝窦高凝和肝损伤明显减弱,伴随着血管内皮细胞中CD40和组织因子(TF)上调的完全抑制。通过配体受体分析,在血管内皮细胞中鉴定了CD40-CD40配体相互作用。在人类LSEC中,IFNγ上调CD40表达,并且通过增加的CD40-CD40配体相互作用进一步诱导TF。与这些发现一致,在患有微循环障碍的人ALI患者中,肝CD40表达显着升高。
结论:我们确定了IFNγ-CD40轴作为ALI微循环障碍的分子机制的关键作用。这一发现可能为ALI的发病机理提供新的见解,并可能有助于ALI患者新的治疗策略的出现。
BACKGROUND: Acute liver failure (ALF) is a life-threatening disorder that progresses from self-limiting acute liver injury (ALI). Microcirculatory disturbance characterized by sinusoidal hypercoagulation and subsequent massive hypoxic hepatocyte damage have been proposed to be the mechanism by which ALI deteriorates to ALF; however, the precise molecular pathway of the sinusoidal hypercoagulation remains unknown. Here, we analyzed ALI patients and mice models to uncover the pathogenesis of ALI with microcirculatory disturbance.
METHODS: We conducted a single-center retrospective study for ALI and blood samples and liver tissues were analyzed to evaluate the microcirculatory disturbance in ALI patients (n = 120). Single-cell RNA sequencing analysis (scRNA-seq) was applied to the liver from the concanavalin A (Con A)‑induced mouse model of ALI. Interferon-gamma (IFNγ) and tumor necrosis factor-alpha knockout mice, and primary human liver sinusoidal endothelial cells (LSECs) were used to assess the mechanism of microcirculatory disturbance.
RESULTS: The serum IFNγ concentrations were significantly higher in ALI patients with microcirculatory disturbance than in patients without microcirculatory disturbance, and the IFNγ was upregulated in the Con A mouse model which presented microcirculatory disturbance. Hepatic IFNγ expression was increased as early as 1 hour after Con A treatment prior to sinusoidal hypercoagulation and hypoxic liver damage. scRNA-seq revealed that IFNγ was upregulated in innate lymphoid cells and stimulated hepatic vascular endothelial cells at the early stage of liver injury. In IFNγ knockout mice treated with Con A, the sinusoidal hypercoagulation and liver damage were remarkably attenuated, concomitant with the complete inhibition of CD40 and tissue factor (TF) upregulation in vascular endothelial cells. By ligand-receptor analysis, CD40-CD40 ligand interaction was identified in vascular endothelial cells. In human LSECs, IFNγ upregulated CD40 expression and TF was further induced by increased CD40-CD40 ligand interaction. Consistent with these findings, hepatic CD40 expression was significantly elevated in human ALI patients with microcirculatory disturbance.
CONCLUSIONS: We identified the critical role of the IFNγ-CD40 axis as the molecular mechanism of microcirculatory disturbance in ALI. This finding may provide novel insights into the pathogenesis of ALI and potentially contribute to the emergence of new therapeutic strategies for ALI patients.