Borlak Jürgen, Spanel Reinhard
Hannover Medical School, Centre for Pharmacology and Toxicology, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
Int J Mol Sci. 2025 Jun 19;26(12):5899. doi: 10.3390/ijms26125899.
Diclofenac is an effective medication for pain and inflammation. However, its use has been linked to hepatitis. To gain insight into diclofenac's ability to cause hepatitis, we investigated the regulation of major effectors of the immune system following daily treatment of minipigs at 3 and 15 mg/kg for 28 days. Histopathology evidenced lobular inflammation, and through a combination of immunogenomics and immunopathology, we detected marked innate and adaptive immune responses. We identified 109 significantly regulated genes linked to neutrophil, monocyte, Kupffer cell, and lymphocyte responses and 32 code for cytokine- and interferon-γ-signaling. In support of wound repair, immunopathology evidenced manifest upregulation of macrophage migration inhibitory factor and CD74. Furthermore, the strong expression of IgG and IgM underscored humoral immune responses. Diclofenac caused an activation of the complement system, especially the C1 inhibitor of the classical pathway and C3 with critical functions in liver regeneration. The marked expression of complement factor B and H of the alternate pathway modulated B-cell responses. Likely, the upregulation of factor H protected hepatocytes from injury by limiting complement-mediated damage of inflamed cells. Additionally, diclofenac treatment elicited marked hepatic expression of lysozyme and KLF6. The latter earmarks M1-polarized Kupffer cells. We observed an extraordinary induction of calprotectin/S100A9 and of the monocyte/macrophage CD163 scavenger receptor, and therefore, we detected innate immune sensing of damaged cells. Lastly, we noted an unprecedented induction of the acute phase reactant SAA1 and DEC-205, which recognize apoptotic and necrotic cells. Together, our results offer mechanistic insights into immune-mediated liver injury patterns following diclofenac treatment.
双氯芬酸是一种治疗疼痛和炎症的有效药物。然而，其使用与肝炎有关。为深入了解双氯芬酸引发肝炎的能力，我们对小型猪每日以3毫克/千克和15毫克/千克的剂量进行治疗，持续28天，研究免疫系统主要效应器的调节情况。组织病理学显示有小叶炎症，通过免疫基因组学和免疫病理学相结合的方法，我们检测到明显的先天性和适应性免疫反应。我们鉴定出109个与中性粒细胞、单核细胞、库普弗细胞和淋巴细胞反应相关的显著调控基因，其中32个编码细胞因子和干扰素-γ信号传导。为支持伤口修复，免疫病理学证明巨噬细胞迁移抑制因子和CD74明显上调。此外，IgG和IgM的强烈表达突出了体液免疫反应。双氯芬酸引起补体系统激活，尤其是经典途径的C1抑制剂和在肝脏再生中起关键作用的C3。替代途径的补体因子B和H的显著表达调节B细胞反应。可能是因子H的上调通过限制补体介导的炎症细胞损伤保护肝细胞免受损伤。此外，双氯芬酸治疗引起溶菌酶和KLF6在肝脏中的显著表达。后者标志着M1极化的库普弗细胞。我们观察到钙卫蛋白/S100A9和单核细胞/巨噬细胞CD163清道夫受体的异常诱导，因此，我们检测到对受损细胞的先天性免疫感知。最后，我们注意到急性期反应物SAA1和DEC-205前所未有的诱导，它们识别凋亡和坏死细胞。总之，我们的结果为双氯芬酸治疗后免疫介导的肝损伤模式提供了机制性见解。
