Wang Feixia, Tang Li, Liang Baoyu, Jin Chun, Gao Liyuan, Li Yujia, Li Zhanghao, Shao Jiangjuan, Zhang Zili, Tan Shanzhong, Zhang Feng, Zheng Shizhong
Department of Integrated TCM and Western Medicine, Nanjing Hospital Affiliated to Nanjing University of Chinese Medicine, Nanjing, China.
Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica, School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, China.
Front Pharmacol. 2021 Apr 23;12:658811. doi: 10.3389/fphar.2021.658811. eCollection 2021.
Acute-on-chronic liver failure (ACLF) is described as a characteristic of acute jaundice and coagulation dysfunction. Effective treatments for ACLF are unavailable and hence are urgently required. We aimed to define the effect of Yi-Qi-Jian-Pi Formula (YQJPF) on liver injury and further examine the molecular mechanisms. In this study, we established CCl-, LPS-, and d-galactosamine (D-Gal)-induced ACLF rat models and LPS- and D-Gal-induced hepatocyte injury models . We found that YQJPF significantly ameliorates liver injury and that is associated with the regulation of hepatocyte necroptosis. Specifically, YQJPF decreased expression of receptor-interacting protein kinase 1 (RIPK1), receptor-interacting protein kinase 3 (RIPK3) and pseudokinase mixed lineage kinase domain-like (MLKL) to inhibit the migration of RIPK1 and RIPK3 into necrosome. YQJPF also reduces the expression of inflammatory cytokines IL-6, IL-8, IL-1β, and TNF-α, which were regulated by RIPK3 mediates cell death. RIPK1 depletion was found to enhance the protective effect of YQJPF. Furthermore, we showed that YQJPF significantly downregulates the mitochondrial reactive oxygen species (ROS) production and mitochondrial depolarization, with ROS scavenger, 4-hydroxy-TEMPO treatment recovering impaired RIPK1-mediated necroptosis and reducing the expression of IL-6, IL-8, IL-1β, and TNF-α. In summary, our study revealed the molecular mechanism of protective effect of YQJPF on hepatocyte necroptosis, targeting RIPK1/RIPK3-complex-dependent necroptosis ROS signaling. Overall, our results provided a novel perspective to indicate the positive role of YQJPF in ACLF.
慢加急性肝衰竭（ACLF）被描述为具有急性黄疸和凝血功能障碍的特征。目前尚无针对ACLF的有效治疗方法，因此迫切需要此类方法。我们旨在确定益气健脾方（YQJPF）对肝损伤的影响，并进一步研究其分子机制。在本研究中，我们建立了由四氯化碳、脂多糖（LPS）和D-半乳糖胺（D-Gal）诱导的ACLF大鼠模型以及由LPS和D-Gal诱导的肝细胞损伤模型。我们发现YQJPF能显著改善肝损伤，且这与肝细胞坏死性凋亡的调控有关。具体而言，YQJPF降低了受体相互作用蛋白激酶1（RIPK1）、受体相互作用蛋白激酶3（RIPK3）和假激酶混合谱系激酶结构域样蛋白（MLKL）的表达，以抑制RIPK1和RIPK3向坏死小体的迁移。YQJPF还降低了炎性细胞因子白细胞介素-6（IL-6）、白细胞介素-8（IL-8）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的表达，这些细胞因子是由RIPK3介导的细胞死亡所调控的。研究发现敲除RIPK1可增强YQJPF的保护作用。此外，我们还表明YQJPF能显著下调线粒体活性氧（ROS）的产生和线粒体去极化，用ROS清除剂4-羟基-TEMPO处理可恢复受损的RIPK1介导的坏死性凋亡，并降低IL-6、IL-8、IL-1β和TNF-α的表达。总之，我们的研究揭示了YQJPF对肝细胞坏死性凋亡保护作用的分子机制，其作用靶点为RIPK1/RIPK3复合体依赖性坏死性凋亡和ROS信号通路。总体而言，我们的结果为表明YQJPF在ACLF中的积极作用提供了一个新的视角。
