Natalia Krasteva, M. Shkodrova, M. Keremidarska-Markova, Dilyana Doncheva-Stoimenova, K. Hristova-Panusheva, Milena Mishonova, M. Chichova
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ATPase activity of both, intact and uncoupled by freezing/thawing mitochondria was determined by the measurement of inorganic phosphate (Pi) released from ATP. The generation of hydrogen peroxide (H2O2) after exposure of mitochondria to GO and GO-NH2 particles was determined by a DCFH-D assay. Results: GO and GO-NH2 particles applied at concentrations of 4 and 50 μg/ml did not affect the ATPase activity of intact mitochondria. In contrast, in uncoupled mitochondria, they demonstrated a stimulating effect on ATPase activity. The impact of GO-NH2 was more substantial and concentration-dependent. ROS production was also higher in GO-NH2-treated mitochondria. Conclusion: The present study demonstrated that GO and GO-NH2 particles can exert a cytotoxic effect on mitochondria even after a short-time of exposure to both types of particles.","PeriodicalId":38867,"journal":{"name":"International Journal Bioautomation","volume":"99 4","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Effect of Graphene Oxide and Ammonia-modified Graphene Oxide Particles on ATPase Activity of Rat Liver Mitochondria\",\"authors\":\"Natalia Krasteva, M. Shkodrova, M. Keremidarska-Markova, Dilyana Doncheva-Stoimenova, K. Hristova-Panusheva, Milena Mishonova, M. Chichova\",\"doi\":\"10.7546/ijba.2024.28.1.000957\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Graphene and its derivatives have become promising materials for biomedical applications in the last decade. Before their widespread application, however, evaluating their toxicity and mechanisms underlying interactions with cellular components is imperative. Aims: Assessment of the effect of two graphene derivatives, pristine graphene oxide (GO) and ammonia-modified GO (GO-NH2) particles, on the ATPase activity of rat liver mitochondria and ROS production. Methods: Liver mitochondria were isolated from male albino rats and treated with different concentrations of GO and GO-NH2 particles (4, 10, 25, and 50 μg/ml). ATPase activity of both, intact and uncoupled by freezing/thawing mitochondria was determined by the measurement of inorganic phosphate (Pi) released from ATP. The generation of hydrogen peroxide (H2O2) after exposure of mitochondria to GO and GO-NH2 particles was determined by a DCFH-D assay. Results: GO and GO-NH2 particles applied at concentrations of 4 and 50 μg/ml did not affect the ATPase activity of intact mitochondria. In contrast, in uncoupled mitochondria, they demonstrated a stimulating effect on ATPase activity. The impact of GO-NH2 was more substantial and concentration-dependent. ROS production was also higher in GO-NH2-treated mitochondria. Conclusion: The present study demonstrated that GO and GO-NH2 particles can exert a cytotoxic effect on mitochondria even after a short-time of exposure to both types of particles.\",\"PeriodicalId\":38867,\"journal\":{\"name\":\"International Journal Bioautomation\",\"volume\":\"99 4\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International Journal Bioautomation\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.7546/ijba.2024.28.1.000957\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Agricultural and Biological Sciences\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal Bioautomation","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.7546/ijba.2024.28.1.000957","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Agricultural and Biological Sciences","Score":null,"Total":0}
引用次数: 0
摘要
在过去十年中,石墨烯及其衍生物已成为生物医学应用中大有可为的材料。然而,在广泛应用之前,评估其毒性以及与细胞成分相互作用的机制势在必行。目的:评估两种石墨烯衍生物--原始氧化石墨烯(GO)和氨修饰的 GO(GO-NH2)颗粒--对大鼠肝脏线粒体 ATP 酶活性和 ROS 生成的影响。研究方法从雄性白化大鼠体内分离出肝脏线粒体,用不同浓度的 GO 和 GO-NH2 粒子(4、10、25 和 50 μg/ml)进行处理。通过测量 ATP 释放的无机磷酸(Pi),测定了完整线粒体和经冷冻/解冻解除耦合的线粒体的 ATP 酶活性。线粒体暴露于 GO 和 GO-NH2 粒子后产生的过氧化氢(H2O2)是通过 DCFH-D 试验测定的。结果显示浓度为 4 和 50 μg/ml 的 GO 和 GO-NH2 粒子不会影响完整线粒体的 ATPase 活性。相反,在非偶联线粒体中,它们对 ATPase 活性有刺激作用。GO-NH2 的影响更大,而且与浓度有关。经 GO-NH2 处理的线粒体产生的 ROS 也更高。结论本研究表明,即使短时间接触 GO 和 GO-NH2 颗粒,它们也能对线粒体产生细胞毒性作用。
Effect of Graphene Oxide and Ammonia-modified Graphene Oxide Particles on ATPase Activity of Rat Liver Mitochondria
Graphene and its derivatives have become promising materials for biomedical applications in the last decade. Before their widespread application, however, evaluating their toxicity and mechanisms underlying interactions with cellular components is imperative. Aims: Assessment of the effect of two graphene derivatives, pristine graphene oxide (GO) and ammonia-modified GO (GO-NH2) particles, on the ATPase activity of rat liver mitochondria and ROS production. Methods: Liver mitochondria were isolated from male albino rats and treated with different concentrations of GO and GO-NH2 particles (4, 10, 25, and 50 μg/ml). ATPase activity of both, intact and uncoupled by freezing/thawing mitochondria was determined by the measurement of inorganic phosphate (Pi) released from ATP. The generation of hydrogen peroxide (H2O2) after exposure of mitochondria to GO and GO-NH2 particles was determined by a DCFH-D assay. Results: GO and GO-NH2 particles applied at concentrations of 4 and 50 μg/ml did not affect the ATPase activity of intact mitochondria. In contrast, in uncoupled mitochondria, they demonstrated a stimulating effect on ATPase activity. The impact of GO-NH2 was more substantial and concentration-dependent. ROS production was also higher in GO-NH2-treated mitochondria. Conclusion: The present study demonstrated that GO and GO-NH2 particles can exert a cytotoxic effect on mitochondria even after a short-time of exposure to both types of particles.