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从类过氧化物酶活性角度评估氧化石墨烯的体外细胞毒性

In vitro cytotoxicity evaluation of graphene oxide from the peroxidase-like activity perspective.

作者信息

Zhang Wei, Sun Ying, Lou Zhichao, Song Lina, Wu Yang, Gu Ning, Zhang Yu

机构信息

State Key Laboratory of Bioelectronics, Jiangsu Key Laboratory for Biomaterials and Devices, School of Biological Science and Medical Engineering & Collaborative Innovation Center of Suzhou Nano Science and Technology, Southeast University, Nanjing 210096, PR China.

Patent Examination Cooperation Jiangsu Centre of the Patent Office, SIPO, Suzhou 215163, PR China.

出版信息

Colloids Surf B Biointerfaces. 2017 Mar 1;151:215-223. doi: 10.1016/j.colsurfb.2016.12.025. Epub 2016 Dec 19.

Abstract

In this study, PEGylated graphene oxide (PEG-GO)-hemin composite structure was constructed. Hemin in the form of nanoscaled aggregates were immobilized on PEG-GO sheets by the π-π stacking super-molecular interaction. Via catalyzing the oxidation of chromogenic substrates, we elicited the obtained PEG-GO-Hemin composite sheets have much higher peroxidase-like activity compared to hemin or PEG-GO alone, which is due to the introduction of enzyme active center of hemin with high dispersity, the excellent affinity to organic substrate through π-π stacking and/or electrostatic adsorption and the rapid electron transfer capability of PEG-GO. Similarly, PEG-GO-Hemin was found to be able to catalyze the oxidation of low density lipoprotein (LDL) by HO, resulting in toxicity to porcine iliac endothelial cells (PIECs) in vitro. Furthermore, we also demonstrated that PEG-GO sheets showed enhanced peroxidase activity when met hemin containing proteins including hemoglobin and cytochrome c. High glucose level (HG) in human umbilical vein endothelial cells (HUVECs) can induce cytochrome c to release from the respiratory chain, thus applying PEG-GO under HG condition could cause a much higher peroxidase-like activity, resulting in the production of hydroxyl radical (OH) and cytochrome c radical (cytochrome c), which eventually enhance the apoptosis. These results suggest GO has potential hazard for biomedical applications in some pathophysiological conditions.

摘要

在本研究中,构建了聚乙二醇化氧化石墨烯(PEG-GO)-血红素复合结构。纳米级聚集体形式的血红素通过π-π堆积超分子相互作用固定在PEG-GO片材上。通过催化显色底物的氧化,我们发现所制备的PEG-GO-血红素复合片材与单独的血红素或PEG-GO相比具有更高的过氧化物酶样活性,这是由于引入了具有高分散性的血红素酶活性中心、通过π-π堆积和/或静电吸附对有机底物具有优异的亲和力以及PEG-GO的快速电子转移能力。同样,发现PEG-GO-血红素能够催化HO对低密度脂蛋白(LDL)的氧化,在体外对猪髂内皮细胞(PIECs)产生毒性。此外,我们还证明,当PEG-GO片材与含血红素的蛋白质(包括血红蛋白和细胞色素c)相遇时,其过氧化物酶活性增强。人脐静脉内皮细胞(HUVECs)中的高血糖水平(HG)可诱导细胞色素c从呼吸链释放,因此在HG条件下应用PEG-GO可导致更高的过氧化物酶样活性,从而产生羟基自由基(OH)和细胞色素c自由基(细胞色素c),最终增强细胞凋亡。这些结果表明,在某些病理生理条件下,氧化石墨烯在生物医学应用中具有潜在危害。

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