Leflunomide-induced cardiac injury in adult male mice and bioinformatic approach identifying Nrf2/NF-κb signaling interplay

Abstract

Leflunomide (LFND) is an immunosuppressive and immunomodulatory disease-modifying antirheumatic drug (DMARD) that was approved for treating rheumatoid arthritis. LFND-induced cardiotoxicity was not fully investigated since its approval. We investigated the cardiac injury in male mice and identified the role of nuclear factor erythroid 2-related factor 2/nuclear factor-κ B (Nrf2/NF-κB) signaling. Male albino mice were assigned into five groups as control, vehicle, and LFND (2.5, 5, and 10 mg/kg). We investigated cardiac enzymes, histopathology, and the mRNA expression of Nrf2, NF-κB, BAX, and tumor necrosis factor-α (TNF-α). The bioinformatic study identified the interaction between LFND and Nrf2/NF-κB signaling; this was confirmed by amelioration in mRNA expression (0.5- to 0.34-fold decrease in Nrf2 and 2.6- to 4.61-fold increases in NF-κB genes) and increased (1.76- and 2.625-fold) serum creatine kinase (CK) and 1.38- and 2.33-fold increases in creatine kinase-MB (CK-MB). Histopathological results confirmed the dose-dependent effects of LFND on cardiac muscle structure in the form of cytoplasmic, nuclear, and vascular changes in addition to increased collagen deposits and apoptosis which were increased compared to controls especially with LFND 10 mg/kg. The current study elicits the dose-dependent cardiac injury induced by LFND administration and highlights, for the first time, dysregulation in Nrf2/NF-κB signaling.

Keywords:

• Cardiac injury
• histopathology
• leflunomide
• mouse
• Nrf2/NF-κB signaling

Acknowledgements

Declaration of Generative AI and AI-assisted technologies in the writing process: During the preparation of this work, authors did not use generative AI and AI-assisted technologies in the writing process.

Author contributions

Conceptualization: AH, MZS, TMA, ASA, and SAZ. Methodology: AAR, LME, and SAZ. Investigation: AAR, AH, AAA, SYA, and SAZ. Software: AAR, AH, HWZK, AAA, and SAZ. Resources: AAR, AH, LME, MZS, TMA, ASA, HWZK, AHE, RA, AAA, and SAZ. Funding acquisition: SYA. Visualization: AAR, AH, LME, MZS, TMA, ASA, HWZK, AHE, RA, and SYA. Validation: AHE and RA. Writing the original draft: AAR, AH, LME, MZS, TMA, ASA, and AHE. Writing, review, and editing: HWZK, RA, AAA, SYA, and SAZ.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

Data are available from the author upon request.

Additional information

Funding

This research was funded by Princess Nourah bint Abdulrahman University Researchers Supporting Project Number (PNURSP2024R713).