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巯基化石墨烯量子点调控巨噬细胞中 ROS 的研究进展。

Development of Thiolated-Graphene Quantum Dots for Regulation of ROS in macrophages.

机构信息

Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri, 64108, USA.

出版信息

Pharm Res. 2016 Nov;33(11):2736-47. doi: 10.1007/s11095-016-2000-7. Epub 2016 Jul 21.

Abstract

PURPOSE

The current study was designed to develop thiolated-graphene quantum dots (SH-GQDs) as a theranostic nanocarrier and evaluate its potential for the optimal scavenging of reactive oxygen species (ROS) in macrophages.

METHODS

SH-GQDs were prepared by hydrothermal pyrolysis of carbon source (citric acid) in the presence of reduced-glutathione (GSH). Raw264.7 cells were treated with varying concentrations of oxLDL (0.5, 1 and 2 μg/ml) in the presence or absence of SH-GQDs and cells were stained with peroxide-sensitive fluorescent dye (DCFDA). Flow cytometry analysis was performed to investigate the expression of MSR and ATP-binding cassette transporter (ABCA1) after such treatments as the negative control, oxLDL treatment and oxLDL treatment in the presence of either GQDs or SH-GQDs.

RESULTS

SH-GQDs had a size ranging from 10 to 30 nm with an average size of 21.3 ± 5.2 nm. The elemental analysis indicated that SH-GQDs are mainly composed of carbon, nitrogen, oxygen and sulfur. The expression levels of ABCA1 in macrophages treated with either LDL or oxLDL were lower than those treated with the media control (the negative control: 100 ± 7.6%; LDL: 82.7 ± 1.2%; and oxLDL: 79.2 ± 1.7%). The level of ABCA1 expression increased as cells were incubated with SH-GQDs (SH-GQDs: 101.5 ± 3.1%). The level of MSR on the surface of macrophages upon being treated with SH-GQDs was lower than those with oxLDL (oxLDL: 112.1 ± 8.8% and SH-GQDs: 91.5 ± 4.2%).

CONCLUSION

The enhancement of lipid efflux and down-regulation of MSR in macrophages by SH-GQDs supported its promising usage as a theranostic nanocarrier to prevent foam cell formation and plaque development.

摘要

目的

本研究旨在开发巯基化石墨烯量子点(SH-GQDs)作为治疗学纳米载体,并评估其在巨噬细胞中最佳清除活性氧(ROS)的潜力。

方法

通过在碳源(柠檬酸)存在下的水热热解制备 SH-GQDs,同时存在还原型谷胱甘肽(GSH)。用不同浓度的 oxLDL(0.5、1 和 2μg/ml)处理 Raw264.7 细胞,存在或不存在 SH-GQDs,并使用过氧化物敏感荧光染料(DCFDA)对细胞进行染色。流式细胞术分析用于研究在阴性对照、oxLDL 处理以及 oxLDL 处理存在 GQDs 或 SH-GQDs 的情况下,这些处理后 MSR 和三磷酸腺苷结合盒转运蛋白(ABCA1)的表达。

结果

SH-GQDs 的尺寸范围为 10 至 30nm,平均尺寸为 21.3±5.2nm。元素分析表明,SH-GQDs 主要由碳、氮、氧和硫组成。用 LDL 或 oxLDL 处理的巨噬细胞中 ABCA1 的表达水平低于用培养基对照处理的细胞(阴性对照:100±7.6%;LDL:82.7±1.2%;oxLDL:79.2±1.7%)。随着细胞孵育 SH-GQDs,ABCA1 的表达水平增加(SH-GQDs:101.5±3.1%)。用 SH-GQDs 处理后,巨噬细胞表面 MSR 的水平低于 oxLDL(oxLDL:112.1±8.8%和 SH-GQDs:91.5±4.2%)。

结论

SH-GQDs 增强了巨噬细胞中的脂质外排和 MSR 的下调,支持其作为治疗学纳米载体的有前途的用途,以防止泡沫细胞形成和斑块发展。

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