Combined effects of IRAK inhibition and pioglitazone on hepatic inflammation and apoptosis in a mouse model of MASLD.

Abstract

Background: Inflammation and apoptosis play crucial role in the progression of liver diseases. This study aimed to evaluate the effects of Interleukin-1 receptor-associated kinase inhibitor (IRAKi) in combination with pioglitazone on the expression of inflammatory and apoptotic markers in the liver of C57BL/6J mice subjected to a high-fat diet (HFD).

Methods: Male C57BL/6J mice were divided into five groups: Control (normal diet, ND), HFD, HFD-IRAKi, HFD-pioglitazone (HFD-PIO), and HFD-IRAKi-PIO. All groups, except ND, were administered HFD for 12 weeks. Subsequently, IRAKi (2 mg/kg, intraperitoneally, three times per week) and pioglitazone (10 mg/kg, orally, daily) were administered for 14 days. Gene and protein expression levels were assessed using real-time PCR and western blot analysis.

Results: Separate administration of IRAKi and pioglitazone significantly reduced the mRNA levels of inflammatory markers IL-1β, IL-6, TNF-α, and NF-κB (p < 0.001). Combined treatment with IRAKi and pioglitazone significantly reduced hepatic triglyceride (TG) and cholesterol content (p < 0.05) and attenuated expression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, NF-κB) and apoptotic markers (cleaved-CASPASE3, Bax), while enhancing Il10 expression. These findings support the therapeutic potential of this combination in metabolic liver diseases such as MASLD.

Conclusion: Co-administration of IRAKi and PIO attenuated markers of inflammation and apoptosis. These findings highlight the potential of IRAKi and pioglitazone as therapeutic agents for metabolic and inflammatory liver diseases, warranting further clinical evaluation.