基于双重类酶活性钯负载氧化石墨烯纳米酶对脑胶质细胞凋亡的影响  

Effect of palladium-supported graphene oxide nanozymes with dual enzyme-like activity on apoptosis of brain glioma cells

作　　者：肖洁 梁超怡[1,2] 张李泽艾 刘斯佳 Xiao Jie;Liang Chaoyi;Zhang-Li Zeai;Liu Sijia(School of Basic Medical Sciences,Guangxi Colleges and Universities Key Laboratory of Biological Molecular Medicine Research,Department of Biochemistry and Molecular Biology,Guangxi Medical University,Nanning 530021,China;Collaborative Innovation Centre of Regenerative Medicine and Medical BioResource Development and Application Co-constructed by the Province and Ministry,Guangxi Key Laboratory of Regenerative Medicine&Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学基础医学院生物化学与分子生物学教研室、广西高校生物分子医学研究重点实验室,南宁市530021 [2]广西医科大学省部共建再生医学与医用生物资源开发与应用协同创新中心、广西再生医学重点实验室、长寿与衰老相关疾病教育部重点实验室,南宁530021

出　　处：《广西医科大学学报》2023年第4期567-574,共8页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81860256);霍英东教育基金会高等院校青年教师资助项目(No.171098);再生医学与医用生物资源开发应用省部共建协同创新中心2023年度重点协同任务(No.CICRMMBR-SR-2023004);广西研究生教育创新计划项目(No.YCSW2022200)。

摘　　要：目的:构建钯负载氧化石墨(GO@Pd)纳米酶,探究其类酶活性对脑胶质细胞凋亡的影响。方法:通过原位生长法合成GO@Pd纳米酶并进行表征;体外检测GO@Pd的类超氧化物歧化酶(SOD)活性、类过氧化物酶(POD)活性和消耗谷胱甘肽(GSH)的能力;并利用亚甲基蓝检测其羟基自由基(·OH^(-))的生成;通过细胞增殖试剂盒评价GO@Pd的体外生物安全性;分别将Pd、GO和GO@Pd与人脑星形胶质母细胞瘤(U-118MG)细胞共同孵育,利用DCFH-DA荧光探针检测各组细胞内活性氧(ROS)水平;通过线粒体膜电位染色检测各组细胞内线粒体膜电位变化。结果:经一系列材料表征表明成功制备了大小均一、分散性良好的GO@Pd纳米酶;类酶活性检测结果显示,GO@Pd同时拥有类SOD和类POD双重类酶活性,并且可以催化H_(2)O_(2)生成·OH^(-)和消耗GSH;当GO@Pd浓度在0~100μg/mL之间,C8-D1A细胞的活率保持在90%以上;与空白对照组相比,GO@Pd组的U-118MG细胞内ROS水平显著升高(P<0.001),并且其线粒体膜电位染色结果红色/绿色荧光强度比值显著降低(P<0.0001)。结论:成功制备了具有双重酶样活性的GO@Pd纳米酶,且其可以通过促进胶质瘤细胞内ROS生成,诱导线粒体损伤,从而引发胶质瘤细胞凋亡。Objective:To construct palladium-supported graphene oxide(GO@Pd)nanozymes,and to explore the effect of their enzyme-like activity on the apoptosis of brain glioma cells.Methods:GO@Pd nanozymes were synthesized and characterized by in-situ growth method.The superoxide dismutase(SOD)-like activity,peroxidase(POD)-like activity,and glutathione(GSH)consumption ability of GO@Pd were detected in vitro.The formation of hydroxyl radical(·OH^(-))was detected by methylene blue.The cell counting kit-8 was used to assess the biosafety of GO@Pd in vitro.Pd,GO and GO@Pd were incubated with human brain astroglioblastoma(U-118MG)cells,respectively,and the intracellular reactive oxygen species(ROS)levels were measured by DCFH-DA fluorescent probes.Mitochondrial membrane potential was detected by mitochondrial membrane potential staining.Results:A series of material characterization showed that GO@Pd nanozymes with uniform size and good dispersion were prepared successfully.The test results of enzyme-like activity showed that GO@Pd possessed both SOD-like and POD-like enzyme activities and could catalyze H_(2)O_(2) to produce·OH^(-)and consume GSH.The viability of C8-D1A cells was maintained above 90% when the concentration of GO@Pd was from 0 to 100μg/mL.Compared with the blank control group,the ROS level of U-118MG cells in GO@Pd group significantly increased(P<0.001),and the red/green fluorescence intensity ratio of mitochondrial membrane potential staining significantly decreased(P<0.0001).Conclusion:The GO@Pd nanozymes with dual enzyme-like activity have been successfully prepared.They can induce mitochondrial damage by promoting intracellular ROS production in glioma cells,thereby triggering apoptosis in glioma cells.

关 键 词：钯负载氧化石墨烯 纳米酶 脑胶质瘤 氧化应激 

分 类 号：R739.41[医药卫生—肿瘤]