Han Mingying, Li Shouzhou, Li Lanrong
Pediatric Intensive Care Unit, Linyi People's Hospital, Linyi, China.
Nutrition Department, Chinese Medicine Hospital in Linyi City, Linyi, China.
J Cell Mol Med. 2021 May 24;25(13):5963-75. doi: 10.1111/jcmm.16357.
Acute liver failure (ALF) is a rare disease characterized by the sudden onset of serious hepatic injury, as manifested by a profound liver dysfunction and hepatic encephalopathy in patients without prior liver disease. In this paper, we aim to investigate whether verapamil, an antagonist of TXNIP, inhibits early ALF through suppressing the NLRP3 inflammasome pathway. Firstly, an ALF mouse model was induced by lipopolysaccharide (LPS)/D-galactosamine (GalN) treatment. The optimal concentration of verapamil in treating early ALF mice was determined followed by investigation on its mechanism in LPS/GalN-induced liver injury. Western blot analysis and co-immunoprecipitation were performed to determine the activation of the TXNIP/NLRP3 inflammasome pathway. Subsequently, overexpression of NLRP3 in mouse liver was induced by transfection with AAV-NRLP3 in vivo and in vitro to identity whether verapamil inhibited early ALF through suppressing the activation of NLRP3 inflammasome. We found that ALF was induced by LPS/GalN in mice but was alleviated by verapamil through a mechanism that correlated with suppression of the NLRP3 inflammasome pathway. Oxidative stress and inflammatory response were induced by intraperitoneal injection of LPS/GalN, but alleviated with injection of verapamil. Overexpression of NLRP3 via AAV in mouse liver in vivo and in vitro reduced the therapeutic effect of verapamil on LPS/GalN-induced ALF. Taken together, the TXNIP antagonist verapamil could inhibit activation of the NLRP3 inflammasome, inflammatory responses and oxidative stress to alleviate LPS/GalN-induced ALF.
急性肝衰竭（ALF）是一种罕见疾病，其特征为严重肝损伤的突然发作，表现为既往无肝病的患者出现严重肝功能障碍和肝性脑病。在本文中，我们旨在研究硫氧还蛋白相互作用蛋白（TXNIP）的拮抗剂维拉帕米是否通过抑制NLRP3炎性小体途径来抑制早期急性肝衰竭。首先，通过脂多糖（LPS）/D-氨基半乳糖（GalN）处理诱导建立急性肝衰竭小鼠模型。确定维拉帕米治疗早期急性肝衰竭小鼠的最佳浓度，随后研究其在LPS/GalN诱导的肝损伤中的作用机制。进行蛋白质免疫印迹分析和免疫共沉淀以确定TXNIP/NLRP3炎性小体途径的激活情况。随后，通过在体内和体外转染腺相关病毒-NLRP3（AAV-NRLP3）诱导小鼠肝脏中NLRP3的过表达，以确定维拉帕米是否通过抑制NLRP3炎性小体的激活来抑制早期急性肝衰竭。我们发现LPS/GalN可诱导小鼠发生急性肝衰竭，但维拉帕米可通过与抑制NLRP3炎性小体途径相关的机制减轻该病症。腹腔注射LPS/GalN可诱导氧化应激和炎症反应，但注射维拉帕米可使其减轻。通过腺相关病毒在体内和体外使小鼠肝脏中NLRP3过表达降低了维拉帕米对LPS/GalN诱导的急性肝衰竭的治疗效果。综上所述，TXNIP拮抗剂维拉帕米可抑制NLRP3炎性小体的激活、炎症反应和氧化应激，从而减轻LPS/GalN诱导的急性肝衰竭。
