Ultrasound-assisted glycyrrhetinic acid/nitric oxide co-loaded albumin nanoparticles for the synergistic treatment of hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is widely recognized as one of the three leading causes of cancer-related death worldwide. Glycyrrhetinic acid (GA) exhibits potent anti-HCC activity, but its poor aqueous solubility limits its clinical application. At high concentrations, nitric oxide (NO) exerts cytotoxic effects on tumor cells. In this study, an ultrasound-assisted albumin nanoparticle system co-loaded with GA and NO (SNO-HSA-GA) was developed for synergistic HCC therapy. The optimized formulation exhibited an average particle size of 232.4 ± 6.2 nm, a zeta potential of -31.2 ± 1.5 mV, and a GA loading of 11.86 ± 1.05%. In vitro drug release exhibited a pH-dependent pattern that was enhanced by ultrasound stimulation. In biological assays, SNO-HSA-GA significantly inhibited hepatoma cell viability, migration, and colony formation while exhibiting minimal toxicity toward normal hepatocytes. Mechanistic investigations indicated that ultrasound-triggered NO release activated the p53/p21 signaling axis, downregulated the expression of CDK4, CDK6, and cyclin D1, and induced cell-cycle arrest in the G 0 /G 1 phase. In vivo experiments demonstrated efficient tumor accumulation, local NO release, substantial inhibition of tumor growth, and increased apoptosis without systemic toxicity following the combined administration of SNO-HSA-GA and ultrasound irradiation. SNO-HSA-GA represents a promising nanoplatform for synergistic HCC therapy.