In recent years, the misuse of acetaminophen (APAP), whether accidental or intentional, has led to a significant increase in cases of acute liver failure worldwide [1,2]. This is primarily due to the formation of a reactive metabolite called N - acetyl - p - benzoquinone imine (NAPQI) [3]. Under normal conditions, NAPQI is detoxified through conjugation with glutathione (GSH) [4]. However, in cases of APAP overdose, GSH reserves are rapidly depleted, leading to the accumulation of NAPQI. This accumulation triggers hepatocyte damage, apoptosis, and subsequent inflammation [5]. Inflammatory responses play a pivotal role in the progression of APAP - induced liver injury [6]. In particular, neutrophils and macrophages are recruited to the liver, where they release pro - inflammatory cytokines such as TNF - α, IL - 6, and IL - 1β, thereby exacerbating liver cell damage and apoptosis [[7], [8], [9]]. Recently, TBXAS1 has attracted interest as a crucial regulator of inflammation [10]. Recently, TBXAS1 has emerged as a key regulator of inflammatory processes [10]. As an enzyme, TBXAS1 catalyzes the conversion of prostaglandin H2 to thromboxane A2 (TXA2), a potent vasoconstrictor and platelet aggregator [11]. TXA2 plays a crucial role in inflammatory activities by promoting the adhesion, migration, and activation of inflammatory cells, as well as stimulating the production of pro - inflammatory cytokines. Elevated levels of TXA2 have been observed in various inflammatory conditions, including liver diseases. However, the specific role of TBXAS1 in APAP - induced liver injury remains to be fully elucidated.

Natural bioactive compounds have garnered significant attention for their potential in disease prevention and treatment, owing to their diverse biological activities and relatively low toxicity [12,13]. Daidzein (DAI), a major isoflavone found in soybeans and other legumes, has been reported to possess a wide range of pharmacological properties, including antioxidant, anti - inflammatory, and anticancer activities [[14], [15], [16]]. Previous studies have suggested that DAI may exhibit hepatoprotective effects against various types of hepatic injuries [17,18]. N - acetylcysteine (NAC), an amino acid derivative with antioxidant, detoxifying, and anti - inflammatory properties, is commonly used in liver protection therapies. It works by enhancing glutathione levels in the body, scavenging free radicals, protecting liver cells from oxidative damage, and supporting the liver's detoxification function. Nevertheless, compared to DAI, NAC has certain limitations, such as weaker antioxidant capacity, lower bioavailability, and a narrower scope of application in specific diseases. DAI and its metabolites (e.g., equol) not only demonstrate stronger antioxidant activity but also offer a broader spectrum of health benefits, such as anticancer properties, improved bone health, and reduced risks of cardiovascular and cerebrovascular diseases. However, the molecular mechanisms underlying the potential protective effects of DAI in APAP-induced liver injury have not yet been fully elucidated.

Given the critical role of TBXAS1 in inflammatory processes and the promising potential of DAI as a natural bioactive compound with hepatoprotective properties, this study aims to investigate whether TBXAS1 may serve as a potential target through which DAI exerts its hepatoprotective effects against APAP - induced liver injury. By elucidating the molecular mechanisms of APAP - induced liver injury and exploring the therapeutic potential of DAI targeting TBXAS1, this research seeks to provide new insights and strategies for managing this condition.