Drug-induced liver injury (DILI) is a prevalent liver disease and the leading cause for acute liver failure (ALF) worldwide. Screening of DILI in patients is central to ensure drug safety and improve therapy efficiency. Mounting evidences revealed that peroxynitrite (ONOO⁻) is involved in the DILI process and can be a potential biomarker for DILI. Thus far, there are few two-photon fluorescence probes for ONOO⁻ that can accomplish this challenging task in DILI liver tissues. Hereby, a peroxynitrite activatable two-photon fluorescence probe BN-PN for the imaging of ONOO⁻ in mice liver was elaborately constructed. The probe specifically reacted with peroxynitrite to furnish 140-fold fluorescence increase in vitro, which elucidated a high sensitivity for ONOO⁻. Thus, subtle changes of ONOO⁻ levels in live cells can be sensitively imaged with this probe by two-photon microscopy. The probe also denoted the overproduction of ONOO⁻ in APAP-induced liver injury, and proved that administration with NAC can effectively alleviate DILI and reduce ONOO⁻ production in mouse liver. Further, the probe demonstrated the rapid rise of ONOO⁻ level in the liver of DILI mice administrated with alcohol. This work disclosed the rational construction of a two-photon fluorescence probe-based DILI screening method, which would help the estimation of drug safety and new drug development.

药物性肝损伤（DILI）是一种普遍的肝病，并且是全球急性肝衰竭（ALF）的主要原因。对患者的DILI进行筛查对于确保药物安全和提高治疗效率至关重要。安装证据透露，过氧亚硝酸盐（ONOO ^- ）是参与DILI过程，并且可以是用于DILI潜力生物标志物。迄今为止，很少有双光子荧光探针ONOO ^-可以实现在帝力肝组织这一具有挑战性的任务。由此，一过氧亚硝酸盐活化双光子荧光探针BN-PN为ONOO的成像^-小鼠肝中精心构成。探针与过氧亚硝酸盐特异性反应，可在体外提供140倍的荧光增强，其阐明了ONOO高灵敏度^- 。因此，ONOO的微妙变化^-在活细胞水平可以敏感地用该探针通过双光子显微镜成像。所述探针也表示ONOO的生产过剩^-在APAP诱导的肝损伤，并证明了与NAC给药可以有效缓解帝力和减少ONOO ^-生产小鼠肝脏。此外，探针证实ONOO迅速崛起^-水平帝小鼠的酒精施用肝脏。这项工作揭示了基于双光子荧光探针的DILI筛选方法的合理构建，这将有助于评估药物安全性和新药开发。