巨胞饮内吞途径介导蒲公英汤剂体进入细胞  

Cellular uptake of dandelion decoctosome is mediated by macropinocytosis

作　　者：李之清 王志清 张艳丽[1] 蒋澄宇[1] LI Zhi-qing;WANG Zhi-qing;ZHANG Yan-li;JIANG Cheng-yu(Department of Biochemistry and Molecular Biology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]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生物化学与分子生物学系

出　　处：《基础医学与临床》2019年第7期925-931,共7页Basic and Clinical Medicine

基　　金：国家重点基础研究发展计划(973计划)(2015CB553400);国家自然科学基金(81570077)

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<0.05);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。Objective To explore 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 inhibited dandelion decoctosome entering into cells(P<0.05).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 significantly impeded the cell entry of dandelion decoctosome(P<0.05).Conclusions Dandelion decoctosome get entrance into cells via endocytosis and predominantly via macropinocytosis.

关 键 词：外泌体 汤剂体 内吞 巨胞饮 

分 类 号：Q291[生物学—细胞生物学]