Abstract:
Objective: To evaluate the effect of a gamma-irradiation-synthesized sodium alginate/acrylamide (SA/AAm) nanogel drug delivery formulation on 5-fluorouracil (5-FU)-induced hepatotoxicity. Methods: Network pharmacology analysis was used to identify overlapping 5-FU targets from the GeneCards and Comparative Toxicogenomics Database, followed by GO and KEGG enrichment analyses. For in vivo validation, male Wistar rats were divided into three groups (n = 6/group) and treated for one month with saline (control), free 5-FU (12.5 mg/kg b.wt, i.p., three times/week), or poly(AAm/SA) nanogel-encapsulated 5-FU (12.5 mg/kg b.wt, i.p., three times/week). Serum liver enzymes, inflammatory markers, pro-survival signaling cascades, metabolic enzymes, and epigenetic targets were subsequently quantified. Results: Network pharmacology analysis identified 539 overlapping 5-FU-related targets from the GeneCards and Comparative Toxicogenomics Database, with enrichment predominantly in liver tissue and pathways associated with inflammation, apoptosis, and pro-survival signaling, including NF-κB and PI3K/AKT. In vivo experiments demonstrated that 5-FU loaded nanogels effectively attenuated conventional 5-FU-induced hepatotoxicity. The nanogel formulation minimized liver injury by normalizing serum aspartate aminotransferase and alanine aminotransferase activities. Mechanistically, it suppressed hepatic inflammation, fibrogenic signaling, cellular proliferation, and metabolic reprogramming by downregulating NF-κB, COX-2, TGF-β1, PI3K, AKT, ERK1/2, Ki-67, and PKM2. Additionally, the nanogels restored hepatic METTL14 expression, mitigating 5-FU-induced disruptions in m6A RNA methylation. Conclusions: The nanogel formulation mitigates 5-FU-induced hepatotoxicity by modulating inflammatory, pro-survival, and metabolic stress pathways, highlighting its potential to improve the hepatic safety profile of 5-FU. However, additional studies investigating histopathology, pharmacokinetics, long-term safety, and antitumor efficacy in tumor-bearing models are required before clinical translation can be considered.