Photodynamic therapy (PDT) involves the administration of a tumor-localizing photosensitizer (PS) followed by light irradiation with a specific wavelength in the presence of molecular oxygen, leading to the generation of cytotoxic singlet oxygen causing the irreversible destruction of malignant and nonmalignant diseases. Hemoporfin is a clinical approval PS widely used for port wine stain and skin diseases in China. In order to enhance singlet oxygen generation for Hemoporfin-mediated PDT, liposomes were utilized as the carrier for Hemoporfin to prolong the blood circulation time and enhance the accumulation in vivo. In addition, the L-buthionine sulfoximine (BSO) and catalase (CAT) were loaded within the hydrophilic cores of liposomes to obtain the BSO/CAT@Liposome-Hemoporfin nanoparticles (BCHL-NPs). The enhanced singlet oxygen generation was sucessfully achieved both in vitro and in vivo, which could be mainly attributed to the increase of Hemoporfin and oxygen concentration in the tumor and the depletion of intracellular glutathione. As a result, the enhanced singlet oxygen generation after BCHL-NPs mediated PDT could significantly suppress the tumor growth without additional side effects as comapred to Hemoporfin.