Amygdalin synergizes with the TNF-α monoclonal antibody infliximab to modulate HSP90 and related necro-inflammatory/oxidative stress pathways in a rat model of hepatic I/R.

Hepatic ischemia/reperfusion (I/R) injury remains a major limitation in liver surgery and transplantation. Amygdalin, a natural glycoside, is known for its antioxidant, anti-inflammatory, and hepatoprotective properties, but its combination with conventional agents is not studied. Accordingly, this study explores a combinatorial approach using a small dose of amygdalin (5 mg/kg) and low-dose infliximab (1.5 mg/kg), a tumor necrosis factor (TNF)-α inhibitor, as a pre-treatment to alleviate I/R-induced liver injury. It also investigates the unexamined role of heat shock protein (HSP)-90 in the modulation of necro-inflammatory pathways during hepatic ischemia/reperfusion injury. Rats were divided into six groups: sham, I/R, infliximab (1.5 and 3 mg/kg), amygdalin (5 mg/kg), and a combination of infliximab (1.5 mg/kg) with amygdalin. Treatments were administered intraperitoneally for three days before I/R induction. Both monotherapies and the combination significantly reduced hepatic expression of HSP90, TNF-α, and phosphorylated-mixed lineage kinase domain-like protein (p-MLKL), while restoring oxidative balance as evidenced by modulation of nuclear factor erythroid 2-related factor 2 (Nrf2), malondialdehyde (MDA), and superoxide dismutase (SOD). The combination therapy additionally suppressed nuclear factor kappa B (NF-κB) in a synergistic manner and enhanced Nrf2 expression compared to amygdalin monotherapy. Correlation analysis revealed strong associations between markers of necroinflammation and oxidative stress, reinforcing the mechanistic interplay between these pathways. HSP90 emerged in the current study as an intermediate modulator linking necroinflammation to oxidative responses. Importantly, the combined therapy exerted a synergistic effect by effectively modulating HSP90 and associated signaling cascades, underscoring its potential as a superior therapeutic strategy.