Abstract: Liposomes, as highly efficient bioactive substance delivery systems, are often limited in their applications due to poor stability and insufficient targeting capabilities. In this study, stearamine was employed as a cationic inducer to prepare cationic liposomes (Lip). Utilizing electrostatic deposition technology, hyaluronic acid (HA), fucoidan (Fuc), chondroitin sulfate (CS), and carboxymethyl chitosan (CMCS) were deposited onto the liposome surface via electrostatic deposition. The microstructure, membrane regulation behavior, and response to environmental stress of the hybrid system were characterized and evaluated using techniques such as particle size and zeta potential analysis, transmission electron microscopy, Fourier transform infrared spectroscopy, Raman spectroscopy, and multiple fluorescent probes. Microstructural results indicated that after modification with HA, Fuc, CS, and CMCS, the Zeta potential of the hybrid systems shifted from a positive value (+32.73 mV) to negative values of −30.77, −41.51, −42.06, and −23.6 mV, respectively. Concurrently, the particle size increased significantly from 95.52 nm to 117.5, 102.9, 122.0, and 137.3 nm (P<0.05), exhibiting a distinct core-shell structure. Multispectral membrane regulation results indicated that polysaccharides primarily interacted with stearylamine and phospholipid headgroups via electrostatic forces and hydrogen bonds. This significantly enhanced the lateral and longitudinal order of lipid molecules, thereby increasing membrane rigidity. More importantly, the polysaccharide coating formed more ordered microdomains in the hydrophobic regions of the liposome membrane by immobilizing phospholipid headgroups and compressing the membrane. Stability results under environmental stress indicated that sulfated polysaccharide-modified liposomes (Fuc and CS) exhibited optimal performance in pH tolerance, ionic strength tolerance, and storage stability. However, under oxidative stress, Fuc unexpectedly promoted phospholipid oxidation, yielding the highest levels of conjugated dienes and malondialdehyde, suggesting its unsuitability for lipid surface modification. This study pioneers new avenues for functionalizing anionic polysaccharides and advances the design and construction of biopolymer-liposome hybrid systems, offering novel directions for related research.