Department of Drug Safety Sciences, Division of Clinical Pharmacology, Nagoya University Graduate School of Medicine, Nagoya, Aichi, 466-8550, Japan.
Toxicology. 2020 Oct;443:152560. doi: 10.1016/j.tox.2020.152560. Epub 2020 Aug 11.
Drug-induced liver injury (DILI) is a major safety concern in drug development. Halothane (HAL), an inhaled anesthetic, induces severe and idiosyncratic liver injury. Ryanodine receptors (RyR) are major intracellular calcium release channels found on the plasma membrane of the endoplasmic reticulum (ER). It has been reported that disordered hepatic calcium homeostasis is a feature of HAL-induced liver injury (HILI) in guinea pigs. However, there are no reports on whether RyR could mediate the pathogenesis of HILI. The aim of the present study was to investigate the effect of RyR on HILI. Ryanodine (RYA, RyR agonist, 50 μg/kg, i.p.) was administered to BALB/c female mice 1 h before HAL administration (15 mmol/kg, i.p.), which significantly elevated plasma transaminase levels and induced severe hepatic inflammation and necrosis. In contrast, dantrolene sodium (DAN, RyR antagonist) treatment significantly suppressed HILI in a dose- and time-dependent manner and alleviated liver damage. The number of infiltrated neutrophils in the liver were higher in the group treated with HAL + RYA than in the group treated with HAL alone, while DAN treatment decreased neutrophil infiltration in HILI. The hepatic mRNA levels of proinflammatory cytokines; chemokines; and factors related to danger signals, neutrophils, oxidative and ER stress, pro-apoptosis, and RyR were significantly increased with RYA pretreatment, whereas these levels were decreased with DAN treatment. These results suggest that RYA exacerbates HILI, and DAN exerts a protective effect against HILI. Hence, our study provides a novel insight regarding the effect of RyR in the mechanism underlying HILI.
药物性肝损伤（DILI）是药物开发中的一个主要安全问题。卤烷（HAL），一种吸入性麻醉剂，可引起严重的、特发性的肝损伤。兰尼碱受体（RyR）是内质网（ER）质膜上的主要细胞内钙释放通道。据报道，肝内钙稳态紊乱是豚鼠 HAL 诱导的肝损伤（HILI）的特征。然而，目前尚无关于 RyR 是否可以介导 HILI 发病机制的报道。本研究旨在探讨 RyR 对 HILI 的影响。在 HAL 给药前 1 小时（15mmol/kg，ip）给 BALB/c 雌性小鼠腹腔注射兰尼碱（RYA，RyR 激动剂，50μg/kg），显著升高血浆转氨酶水平，并诱导严重的肝炎症和坏死。相比之下，丹曲林钠（DAN，RyR 拮抗剂）治疗在剂量和时间依赖性方式下显著抑制 HILI，并减轻肝损伤。与仅用 HAL 处理的组相比，用 HAL+RYA 处理的组中肝内浸润的中性粒细胞数量更高，而 DAN 处理减少了 HILI 中的中性粒细胞浸润。用 RYA 预处理可显著增加肝内促炎细胞因子、趋化因子和与危险信号、中性粒细胞、氧化和 ER 应激、促凋亡和 RyR 相关的因子的 mRNA 水平，而 DAN 治疗则降低了这些水平。这些结果表明，RYA 加重了 HILI，而 DAN 对 HILI 具有保护作用。因此，我们的研究为 RyR 在 HILI 发病机制中的作用提供了新的见解。
