Fu Juan, Huang Qiuhong, Sun Changfeng, Li Shuyi, Wang Qingsong, Sheng Yunjian, He Binfeng, You Zaichun
Department of General Medicine, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China.
Department of Infectious Diseases, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, China.
Sci Rep. 2025 Jul 21;15(1):26395. doi: 10.1038/s41598-025-10764-x.
Acetaminophen (APAP) overdose is a common cause of drug-induced liver injury (DILI), which can lead to sterile inflammation and progress to acute liver failure and even death. However, there are currently limited therapeutic options available. Iinterleukin-37 (IL-37) is considered as an anti-inflammatory cytokine. The role and novel mechanism of IL-37 on DILI are still unknown. Male C57BL/6 mice were pretreated with IL-37 for 2 h prior to intraperitoneal injection of acetaminophen (APAP). Hepatic function was assessed by measuring serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH). Liver tissue damage was evaluated via hematoxylin and eosin (H&E) staining. The inflammatory response was characterized by immunohistochemical (IHC) analysis of myeloperoxidase (MPO) and lymphocyte antigen 6 complex locus G (Ly6G), and the levels of interleukin-6 (IL-6), IL-10, transforming growth factor-beta 1 (TGF-β1), and tumor necrosis factor-alpha (TNF-α) in liver tissue. Oxidative stress status was determined by measuring superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) levels. CYP2E1 mRNA expression was analyzed using qPCR. Protein expression of phosphorylated p38 (pP38), phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2), and phosphorylated p65 (pP65) was evaluated by Western blotting. Compared to the model group without recombinant human IL-37 treatment, the model + IL-37 group exhibited significantly attenuated liver injury, characterized by reduced neutrophil infiltration, decreased levels of pro-inflammatory mediators IL-6 and TNF-α, and elevated levels of the anti-inflammatory cytokines IL-10 and TGF-β1. The levels of pp38, pERK, and pp65 in liver tissue were significantly suppressed in the Model + IL-37 group compared to the Model group at 24 h. Furthermore, MDA levels were significantly lower in the IL-37-treated group relative to the model group, while SOD activity showed no significant difference. Our results also indicate that neither CYP2E1 mRNA relative expression nor GSH levels differed significantly between the model group and the IL-37-treated group at either 4 h or 24 h after APAP exposure. IL-37 has a significant protective effect against acetaminophen-induced liver injury (AILI) by suppressing the inflammatory response involved in the MAPK-NF-κB/p65 signalling pathway. Our study suggests IL-37 as a potential therapeutic strategy for DILI.
对乙酰氨基酚（APAP）过量是药物性肝损伤（DILI）的常见原因，可导致无菌性炎症，并进展为急性肝衰竭甚至死亡。然而，目前可用的治疗选择有限。白细胞介素-37（IL-37）被认为是一种抗炎细胞因子。IL-37在DILI中的作用和新机制仍不清楚。雄性C57BL/6小鼠在腹腔注射对乙酰氨基酚（APAP）前2小时用IL-37预处理。通过测量血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）和乳酸脱氢酶（LDH）水平评估肝功能。通过苏木精和伊红（H&E）染色评估肝组织损伤。通过免疫组织化学（IHC）分析髓过氧化物酶（MPO）和淋巴细胞抗原6复合体基因座G（Ly6G）以及肝组织中白细胞介素-6（IL-6）、IL-10、转化生长因子-β1（TGF-β1）和肿瘤坏死因子-α（TNF-α）的水平来表征炎症反应。通过测量超氧化物歧化酶（SOD）、丙二醛（MDA）和谷胱甘肽（GSH）水平来确定氧化应激状态。使用qPCR分析CYP2E1 mRNA表达。通过蛋白质印迹法评估磷酸化p38（pP38）、磷酸化细胞外信号调节激酶1/2（pERK1/2）和磷酸化p65（pP65）的蛋白质表达。与未用重组人IL-37治疗的模型组相比，模型+IL-37组肝损伤明显减轻，其特征为中性粒细胞浸润减少、促炎介质IL-6和TNF-α水平降低以及抗炎细胞因子IL-10和TGF-β1水平升高。与模型组相比，模型+IL-37组在24小时时肝组织中pp38、pERK和pp65的水平明显受到抑制。此外，IL-37治疗组的MDA水平相对于模型组显著降低，而SOD活性无显著差异。我们的结果还表明，在APAP暴露后4小时或24小时，模型组和IL-37治疗组之间CYP2E1 mRNA相对表达和GSH水平均无显著差异。IL-37通过抑制MAPK-NF-κB/p65信号通路中的炎症反应，对乙酰氨基酚诱导的肝损伤（AILI）具有显著的保护作用。我们的研究表明IL-37是DILI的一种潜在治疗策略。
