Micro- and nanoplastics (MNPs) form protein corona (PC) upon contact with biological fluids, but their impact on the intracellular transport, distribution, and toxicity of MNPs remains unclear. Fetal bovine serum (FBS) and bovine serum albumin (BSA) were used to simulate in vivo environment, this study explored their influence on the transport and toxicity of polystyrene (PS) MNPs in zebrafish liver (ZFL) cells. Results showed PS MNPs were wrapped by proteins into stable complexes. Nanoparticles (NP, 50 nm) and their protein complexes (NP@PC) were internalized by cells within 6 h, with PC formation enhancing NP uptake. NP primarily entered cells through clathrin- and caveolae-mediated endocytosis, while NP@PC via clathrin-mediated pathways. Internalized particles were predominantly in lysosomes where PC degraded and some were also in mitochondria. Eventually, particles were expelled from cells through energy-dependent lysosomal pathways and energy-independent membrane penetration mechanisms. Notably, PC formation limited the clearance of NP. In toxicity, NP had a more severe impact than microplastics (MP, 5 μm). FBS more effectively mitigated PS MNPs-induced reactive oxygen species accumulation, subcellular structural damage, and dysregulation of glycolipid metabolism than BSA did. This study elucidates the modulatory role of PC on biological effects of MNPs, providing safety and risk management strategies.