Targeting AMPK with ivabradine attenuates cyclophosphamide-induced hepatotoxicity: Crosstalk with MAPK, JAK1/STAT3, and PI3K/Akt pathways

Abstract

Aims: This study explores the molecular pathways through which Ivabradine (IVN) exerts protective effects against Cyclophosphamide (CP)-induced hepatotoxicity, aiming to identify the underlying mechanisms involved.

Main methods: Animals were assigned at random into four groups (10 rats in each group): Group 1 was administered 1 mL of distilled water orally for 10 consecutive days, along with a single intraperitoneal injection of 0.9% saline on the seventh day. Group 2 received distilled water for 10 days and a single CP injection (200 mg/kg, IP) on day 7. Groups 3 and 4 were administered IVN at either 5 or 10 mg/kg orally each day for 10 consecutive days, plus CP on day 7. Rats were euthanized for biochemical, histological, immunostaining, qRT-PCR, and western blot assessments.

Key findings: Rats treated with CP exhibited notable elevations in liver enzymes (ALT, AST) along with decreased levels of antioxidant indicators (HO-1, Nrf2, GSH). Concentrations of MPO, MDA, and iNOS, along with IL-1β, TNF-α, and IL-6, were markedly elevated (p < 0.01). Immunostaining showed elevated NF-kB p65 and caspase-9, aligning with observed liver histopathology. Conversely, IVN administration reduced hepatic enzymes and alleviated tissue alterations. It provided antioxidant defense by correcting redox imbalance and lowering inflammation through targeting the p38MAPK/NF-κB p65 axis and the JAK1-STAT3 pathway (p < 0.01). IVN also prevented CP-induced PI3K-Akt suppression and reduced caspase-related apoptotic activity.

Significance: The current findings indicate that IVN could represent a valuable treatment option to mitigate CP-induced liver injury via its antioxidant, anti-inflammatory, and apoptosis- suppressing properties, thereby supporting the need for further exploration in future studies.

Keywords: AMPK; Cyclophosphamide; Hepatoxicity; Ivabradine; MAPK; PI3K.