广西医科大学学报2025，Vol.42Issue(2)：192-201,10.DOI:10.16190/j.cnki.45-1211/r.2025.02.005
铑—铜双金属纳米酶对骨肉瘤细胞凋亡的影响
Effect of rhodium-copper bimetallic nanozymes on apoptosis of osteosarcoma cells
摘要
Abstract
Objective:To construct a graphdiyne-supported rhodium-copper bimetallic(Rh-Cu/GDY)nanozyme system through an innovative preparation process,so as to explore the unique enzyme-like activity of the nano-zymes and their effect on the apoptosis of osteosarcoma(OS)cells.Methods:Synthesis and characterization of Rh-Cu/GDY nanozymes were carried out by the hydrothermal method.The peroxidase-like(POD)activity,catalase-like(CAT)activity of Rh-Cu/GDY,and its ability to consume glutathione(GSH)were accurately de-tected in vitro.Additionally,the abilities of Rh-Cu/GDY to catalyze hydrogen peroxide(H₂O₂)to generate oxygen(O₂)and hydroxyl radicals(·OH)and to consume GSH through its CAT and POD enzyme activities were de-tected using a dissolved oxygen meter,3,3',5,5'-tetramethylbenzidine(TMB),and 5,5'-dithiobis(2-nitrobenzoic acid)(DTNB)fluorescent probes.The cytotoxicity and anti-tumor activity of Rh-Cu/GDY against human umbili-cal vein endothelial cells(HUVECs),OS cells 143B and MG63 were detected by cell counting kit-8(CCK-8).The levels of intracellular reactive oxygen species(ROS)in each group of cells were detected using HPF and[Ru(DPP)3]Cl2 fluorescent probes.The JC-1 mitochondrial fluorescence staining technique was used to evaluate the mitochondrial level of tumor cells.The Ki67 immunofluorescence staining experiment was conducted to detect the expression of Ki67 protein in tumor cells.The western blotting experiment for Bax protein was performed to evaluate the expression of intracellular Bax protein in tumor cells treated with nanozymes at different concentra-tions.Results:Transmission electron microscopy(TEM),Zeta potential analysis,high-resolution transmission electron microscopy(HRTEM),X-ray photoelectron spectroscopy(XPS),and energy-dispersive X-ray spectros-copy mapping(EDS-mapping)confirmed the successful synthesis of the Rh-Cu/GDY nanozymes.The enzyme-like activity assay showed that Rh-Cu/GDY nanozymes could effectively mimic POD and CAT activities while consuming GSH.CCK-8 experimental results confirmed that Rh-Cu/GDY nanozymes had good biocompatibility and anti-tumor ability.Compared with the blank control group,the level of intracellular ROS in the 143B cells of the Rh-Cu/GDY group showed an extremely significant upward trend.Compared with the blank control group,the expression of red fluorescence of JC-1 mitochondria in the Rh-Cu/GDY group was significantly increased.Only weak fluorescence of Ki67 was observed in the Ki67 immunofluorescence assay,and the expression level of Bax protein was also gradually increased with the increase of the concentration of the nanozymes.Conclusion:The Rh-Cu/GDY nanozymes with multiple enzyme-like activities are successfully prepared,and they can achieve the effect of killing OS by promoting the generation of ROS in OS cells and inducing apoptosis.The Rh-Cu/GDY nanozymes are is an anti-tumor nanomaterial with promising prospects.关键词
铑—铜双金属/纳米酶/活性氧/骨肉瘤Key words
rhodium-copper bimetal/nanozymes/reactive oxygen species/osteosarcoma分类
临床医学引用本文复制引用
王韩洁,叶玉婷,广怡然,黄章睿,钟静萍..铑—铜双金属纳米酶对骨肉瘤细胞凋亡的影响[J].广西医科大学学报,2025,42(2):192-201,10.基金项目
国家自然科学基金资助项目(No.82160429) （No.82160429）
