上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2018, Vol. 38 ›› Issue (1): 42-.doi: 10.3969/j.issn.1674-8115.2018.01.008

• 论著（基础研究） • 上一篇    下一篇

球形金纳米颗粒对骨髓间充质干细胞自噬流和外泌体分泌的影响

颜春1，马玉洁2，谢玮岳1，杨庆源3，孙奋勇3，杨志2，薛松1，连锋1   

  1. 1. 上海交通大学 医学院附属仁济医院心外科，上海 200127；2. 上海交通大学 电子信息与电气工程学院薄膜与微细技术教育部重点实验室，上
    海 200240；3. 同济大学 医学院附属第十人民医院检验科，上海 200072
  • 出版日期:2018-01-28 发布日期:2018-03-09
  • 作者简介:颜 春（1990—），男，硕士生；电子信箱：1146911579@qq.com。
  • 基金资助:
    上海交通大学医工交叉研究基金（YG2015MS51）

Effect of gold nanoparticle on autophagy flux and exosome secretion of mesenchymal stem cells

YAN Chun1, MA Yu-jie2, XIE Wei-yue1, YANG Qing-yuan3, SUN Feng-yong3, YANG Zhi2, XUE Song1, LIAN Feng1   

  1. 1. Department of Cardiovascular Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127; 2. Ministry of Education Key Laboratory of Thin Film and Microfabrication Technology, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240; 3. Department of Clinical Laboratory Medicine, Shanghai Tenth People’s Hospital of Tongji University, Shanghai 200072
  • Online:2018-01-28 Published:2018-03-09
  • Supported by:
    Shanghai Jiao Tong University Medicine-Engineering Foundation, YG2015MS51

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摘要： 目的· 探讨球形金纳米颗粒（AuNPs）阻断大鼠骨髓间充质干细胞（MSCs）的自噬流对外泌体分泌的影响。方法· 体外培
养大鼠MSCs，分别以不同浓度AuNPs（0、15、30、90、150 μg/mL）处理细胞，CCK-8 法检测细胞活力，以评估AuNPs 细胞毒
性。细胞分3 组：正常对照组、AuNPs 组（20 μg/mL AuNPs 作用72 h）和阳性对照组（5 μmol/L 氯喹作用4 h）。透射电镜下观察
超微结构，超速离心提取细胞上清液外泌体，行纳米颗粒分析仪和电镜检测。共聚焦显微镜观察细胞自噬，Western blotting 验证。
结果· 0 ～ 30 μg/mL 的AuNPs 对细胞增殖的影响小。AuNPs 组的MSCs 外泌体分泌增多，体积减小，LC3 和p62 表达增高（均
P<0.05），自噬小体增多，溶酶体和自噬小体融合受阻。结论· AuNPs 阻断MSCs 自噬流，调节外泌体分泌。

关键词: 金纳米颗粒, 间充质干细胞, 外泌体, 自噬

Abstract:

Objective · To investigate the effect of gold nanoparticles (AuNPs) on exosome secretion of mesenchymal stem cells (MSCs) via blocking
autophagy flux. Methods · Rat derived MSCs were treated with AuNPs (0, 15, 30, 90, 150 μg/mL), and the cell proliferation was detected with CCK-8
assay to evaluate the cytotoxicity of AuNPs. Then, MSCs were divided into normal control group, AuNPs group (20 μg/mL AuNPs for 72 h) and positive
control group (5 μmol/L chloroquine for 4 h). Transmission electron microscopy was used to observe the ultrastructure of MSCs, and exosomes were
extracted by ultracentrifugation and tested with nanoparticle analyzer and scan electron microscopy. The autophagy was observed by confocal microscopy
and verified by Western blotting. Results · Low concentrations of AuNPs (0 to 30 μg/mL) slightly affected the cell proliferation. MSCs of AuNPs group
secreted more abundant and smaller exosomes than those of control group, and expressed significantly more LC3 and p62 (both P<0.05). Autophagosomes in AuNPs group were increased and fusion of lysosomes and autophagosomes was blocked. Conclusion · AuNPs block the autophagy flux and regulates the secretion of exosomes.

Key words: gold nanoparticle, mesenchymal stem cell, exosome, autophagy