Objective To address the limited oral utilization of metformin hydrochloride (MET) and the insufficient delivery efficiency of conventional liposomes, MET-loaded nanoliposomes modified with Agaricus bisporus chitosan (ACS) (ACS-modified liposomes, A-M-L) were developed, and their physicochemical properties, storage stability, cytocompatibility, and effects on glucose consumption by HepG2 cells were evaluated.Methods ACS was used as a substitute for crustacean chitosan to prepare MET-loaded nanoliposomes using the thin-film dispersion-ultrasonication method. The structural characteristics and microscopic morphology were observed by Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM), and particle size, polydispersity index (PDI), zeta potential, encapsulation efficiency, storage stability, cytotoxicity, and glucose consumption were determined.Results ACS modification improved the physicochemical properties of the liposomes. The 0.5% ACS-modified group exhibited a particle size of approximately 159.45 nm, a low PDI of 0.10, and an initial encapsulation efficiency of 87.46%. Microscopic characterization indicated the successful encapsulation of the drug. After storage at 4 ℃ for 21 days, the particle size increased by only 1.8%, while the encapsulation efficiency remained at 83.1%, indicating good stability under the storage conditions used in this study. In vitro experiments demonstrated that the 1.0% ACS-modified group promoted glucose consumption by HepG2 cells and enhanced glucose metabolism, with glucose consumption reaching 4.9 mmol/L, higher than that in the MET drug group (3.5 mmol/L) and the unmodified liposome group (2.3 mmol/L). In addition, ACS modification effectively attenuated the cytotoxicity of the free drug.Conclusion ACS-modified MET-loaded nanoliposomes can improve the dispersibility, encapsulation efficiency, and storage stability of liposomes and increase glucose consumption by HepG2 cells in vitro.