未折叠蛋白反应
衣霉素
坏死性下垂
塔普斯加尔金
内质网
生物
肝细胞
细胞生物学
活力测定
程序性细胞死亡
药理学
内分泌学
细胞凋亡
生物化学
体外
作者
Rakesh K. Arya,Emily Huang,Megan R. McMullen,Kyle L. Poulsen,Jian Wu,Jared Travers,Evangelia Paouri,Dimitrios Davalos,Laura E. Nagy
标识
DOI:10.1101/2025.09.24.677754
摘要
Background and Aim: Endoplasmic reticulum (ER) stress is an important contributor to liver disease progression, including alcohol-associated liver disease (ALD). While receptor-interacting protein kinase-3 (RIP3) and mixed lineage kinase domain-like pseudokinase (MLKL) are known for their roles in necroptosis, emerging evidence highlights their non-canonical functions in metabolic regulation and cellular stress responses. However, their specific role in regulating hepatic ER stress remains unclear. This study investigates how RIP3, its kinase activity, and MLKL regulate ER stress pathways during chronic ethanol exposure and pharmacological ER stress induction. Methods: Rip3-/-, Rip3K51A/K51A and Mlkl-/- mice alongside WT controls and pharmacological necroptosis inhibitors were used to study the role of RIP3 and MLKL in modulating ER stress. Chronic ethanol feeding and pharmacological agents (tunicamycin, thapsigargin) were utilized to induce ER stress in vivo and in isolated primary hepatocytes. ER stress markers were assessed by qPCR and western blot, ER expansion was evaluated by confocal microscopy, and hepatocyte viability was measured using MTS assay. Results: Chronic ethanol increased expression of ER stress markers in WT mice; this response was attenuated in Rip3-/- mice. Tunicamycin exposure increased hepatic ER stress markers in WT mice; this response was diminished in Rip3-/-, Rip3K51A/K51A and Mlkl-/- mice. In primary hepatocytes, genetic and pharmacological inhibition of RIP3 and MLKL also reduced thapsigargin-induced ER stress responses. Hepatocytes isolated from Rip3-/-, Rip3K51A/K51A and Mlkl-/- mice exhibited enhanced cell viability under ER stress conditions compared to hepatocytes from WT mice, which was associated with ER expansion as a potential mechanism for mitigating ER stress. Conclusion: This study highlights a novel function of RIP3 and MLKL in regulating hepatic ER stress responses, expanding their known roles beyond programmed necrosis.
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