Roles of diclofenac and its metabolites in immune activation associated with acute hepatotoxicity in TgCYP3A4/hPXR-humanized mice
Roles of diclofenac and its metabolites in immune activation associated with acute hepatotoxicity in TgCYP3A4/hPXR-humanized mice
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双氯芬酸及其代谢物在 TgCYP3A4/hPXR 人源化小鼠急性肝毒性相关免疫激活中的作用
DOI：
10.1016/j.intimp.2020.106723
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发表时间：
2020-09-01
影响因子：
5.6
通讯作者：
Dai, Renke
中科院分区：
文献类型：
--
作者：
Jiang, Weifan;Dai, Tianming;Dai, Renke
双氯芬酸(DCF)是一种广泛使用的非类固醇抗炎药，但它具有较高的药物性肝损伤(DILI)风险。尽管存在苯醌-亚胺加合物途径，但免疫毒性最近被认为是DILI的另一个因素。然而，这种免疫反应仍然难以捉摸。在本研究中，分别用DCF和DCF代谢物对TgCYP3A4/hPXR人源化小鼠急性肝毒性模型的免疫应答进行了研究。单次腹腔注射80 mg/kg DCF的药代动力学结果表明，DCF的主要代谢产物为4‘-羟基双氯芬酸(4’-OH-DCF)、5-羟基双氯芬酸(5-OH-DCF)和双氯芬酸葡萄糖醛酸苷(DCF-G)。不仅DCF，而且DCF的代谢产物也可直接引起不同程度的急性肝损伤，在短时间内显著升高TgCYP3A4/hPXR人源化小鼠的血清ALT水平。此外，三种DCF代谢物均能不同程度地直接刺激血清免疫相关因子的显著升高。转录组分析显示，DCF-G处理的小鼠肝脏差异表达基因主要参与“免疫系统过程”和“细胞死亡”，并与“IL-17信号通路”和“肿瘤坏死因子-α信号通路”有关，而5-OH-DCF对这些基因的表达影响不大。这些结果表明，代谢产物DCF-G在激活肝脏免疫系统中起重要作用，可能参与了DCF所致急性肝损伤的发病机制。
Diclofenac (DCF) is a widely used nonsteroidal anti-inflammatory drug, but it comes with a high risk of drug-induced liver injury (DILI). Despite the quinone-imine adduct pathways, the immunotoxicity is recently considered as another factor for DILI. However, such immune responses are still elusive. In the present study, investigation of the immune response in the acute hepatotoxicity model of TgCYP3A4/hPXR-humanized mice was conducted by administration of DCF and DCF metabolites, respectively. In a single dose intraperitoneal injection of 80 mg/kg DCF, the pharmacokinetic results showed the major DCF metabolites, including 4'-hydroxy-diclofenac (4'-OH-DCF), 5-hydroxy-diclofenac (5-OH-DCF) and diclofenac glucuronide (DCF-G) were generated after DCF treatment. Not only DCF, but those DCF metabolites could also directly cause different degrees of acute liver injury as significantly increased the serum ALT levels in a short time period in the TgCYP3A4/hPXR-humanized mice. Furthermore, the three DCF metabolites could directly stimulate the significant elevation of serum immune-related factors in varying degrees. Transcriptome analysis revealed the differentially expressed genes in the liver of DCF-G treated mice were mostly involved with the "immune system process" and "cell death" and related to "IL-17 signaling pathway" and "TNF-alpha signaling pathway", but 5-OH-DCF had little effect on the expressions of those genes. These results indicate that the metabolite DCF-G plays an important role in the activation of the hepatic immune system, which might be involved in the pathogenesis of DCF-induced acute liver injury.