Yi-Qi-Jian-Pi Formula Suppresses RIPK1/RIPK3-Complex-Dependent Necroptosis of Hepatocytes Through ROS Signaling and Attenuates Liver Injury in Vivo and in Vitro

Abstract

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 CCl4-, LPS-, and d-galactosamine (D-Gal)-induced ACLF rat models in vivo and LPS- and D-Gal-induced hepatocyte injury models in vitro. We found that YQJPF significantly ameliorates liver injury in vivo and in vitro 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 via ROS signaling. Overall, our results provided a novel perspective to indicate the positive role of YQJPF in ACLF.

Roles of necroptosis in alcoholic liver disease and hepatic pathogenesis

“… 63 The latest research indicated that Yi‐Qi‐Jian‐Pi Formula (YQJPF) decreased hepatocyte necroptosis in the acute‐on‐chronic liver failure (ACLF) model, consisting of inhibiting the expression of RIPK1 and RIPK3 and blocking the formation of necrosome. 64 Meanwhile, YQJPF reduced the level of ROS in hepatocytes, which had the same effect as selective RIPK1 inhibitor Nec‐1, suggesting that YQJPF inhibited necroptosis by reducing ROS signalling.…”

Cpd-42 protects against calcium oxalate nephrocalcinosis-induced renal injury and inflammation by targeting RIPK3-mediated necroptosis

“…RIPK1, RIPK3 and MLKL are all targets of intervention against necroptosis-mediated inflammation and pathology ( Dannappel et al, 2014 ; Mulay et al, 2016 ; Martens et al, 2020 ; Liu et al, 2022 ). To date, various inhibitors targeting RIPKs have been developed ( Martens et al, 2020 ; Wang et al, 2021 ) and have been shown to protect against the progression of various kidney diseases ( Mulay et al, 2016 ; Liu et al, 2018 ; Wang et al, 2019 ; Liu et al, 2022 ). We previously reported that two RIPK1 inhibitors, Cpd-71 and wogonin, attenuated cisplatin-induced nephropathy and were superior to the classical RIPK1 inhibitor Nec-1.…”

Ancient Herbal Formula Mahuang Lianqiao Chixiaodou Decoction Protects Acute and Acute-on-Chronic Liver Failure via Inhibiting von Willebrand Factor Signaling

“…Therefore, the CCl 4 +LPS/D−GalN-induced ACLF mouse model has also been used to test the potential therapeutic effects of drugs on ACLF [ 39 ]. We replicated murine models of ACLF and ALF with mortality by LPS+D−GalN and CCl 4 , as previously described [ 36 , 40 ]. Compared with ALF mice, latent cirrhotic ACLF mice showed a partially differentiated phenotype, including more serious liver function damage (higher ALT and AST), delayed time to death, and different histological morphology.…”