基础医学与临床 ›› 2019, Vol. 39 ›› Issue (7): 925-931.

• 研究论文 •    下一篇

巨胞饮内吞途径介导蒲公英汤剂体进入细胞

李之清1,王志清1,张艳丽2,蒋澄宇3   

  1. 1. 中国医学科学院 北京协和医学院 基础医学研究所
    2. 中国医学科学院基础医学研究所
    3. 中国医学科学院基础医学研究所 北京协和医学院基础学院
  • 收稿日期:2019-04-08 修回日期:2019-05-21 出版日期:2019-07-05 发布日期:2019-07-02
  • 通讯作者: 张艳丽 E-mail:zhyanli@gmail.com
  • 基金资助:
    国家自然科学基金;国家重点基础研究发展计划(973计划)

Cellular uptake of dandelion decoctosome is mediated by macropinocytosis

  • Received:2019-04-08 Revised:2019-05-21 Online:2019-07-05 Published:2019-07-02
  • Contact: Yan-Li ZHANG E-mail:zhyanli@gmail.com

摘要: 目的 探究细胞摄取蒲公英汤剂体的机制。方法 在人肺腺癌细胞系A549细胞模型上,用内吞通路的特异性化学抑制剂,以及敲减内吞通路中关键蛋白的手段,通过流式细胞计量术和激光共聚焦显微镜检测,评估荧光标记的蒲公英汤剂体进入细胞的机制。结果 亲溶酶体物质能显著抑制蒲公英汤剂体进入细胞(P<0.01);巨胞饮抑制剂细胞松弛素D和阿米洛利能显著降低蒲公英汤剂体的胞内摄入(P<0.001),且呈现剂量依赖性,敲减Rac1和PAK1也有效地抑制其进入细胞(P<0.05)。结论 蒲公英汤剂体通过内吞进入细胞且主要由巨胞饮介导。

关键词: 外泌体, 汤剂体, 内吞, 巨胞饮

Abstract: Objective To characterize the mechanism of cellular uptake of dandelion decoctosome. Methods Internalization of fluorescently-labeled dandelion decoctosome was evaluated in A549 cells. By using a range of chemical inhibitors to block specific pathways as well as RNA interference (RNAi) to knockdown key proteins involved in individual endocytic pathways, cellular uptake of dandelion decoctosome was assessed using flow cytometry and confocal microscopy. Results Lysosomotropic agents significantly decreased dandelion decoctosome entering into cells(P<0.01). Moreover, inhibitors of macropinocytosis, including cytochalasin D and amiloride significantly reduced the internalization of dandelion decoctosome in a dose dependent manner(P<0.001). Consistently, knockdowning of Rac1 and PAK1 by RNAi impeded the cell entry of dandelion decoctosome(P<0.05). Conclusions dandelion decoctosome enters cells via endocytosis and predominantly via macropinocytosis.

Key words: exosomes, decoctosome, endocytosis, macropinocytosis

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