细胞力学作用对MDCK细胞系微组织形态发生的影响  

Effect of cell mechanics on morphogenesis of MDCK lobular organoid

作　　者：曹煜 吴当 欧阳明星 邓林红[1] Cao Yu;Wu Dang;Ouyang Mingxing;Deng Linhong(Institute of Biomedical Engineering and Health Sciences,School of Medical and Health Engineering,2School of Pharmacy&School of Biological and Food Engineering,Changzhou University,Changzhou 213164,Jiangsu Province,China)

机构地区：[1]常州大学医学与健康工程学院生物医学工程与健康科学研究院,江苏省常州市213164 [2]常州大学药学院生物与食品工程学院,江苏省常州市213164

出　　处：《中国组织工程研究》2024年第31期5003-5009,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(NSFC12372312,NSFC11872129),项目负责人:欧阳明星;国家自然科学基金项目(11532003),项目负责人:邓林红。

摘　　要：背景:体内组织器官的发生是一个精确而自主调控的过程,生物力学因素在该宏观尺度上的功能是值得探索的基础科学问题。目的:探讨细胞力学作用对3D Madin-Darby犬肾(MDCK)上皮微组织形态发生的影响。方法:利用荧光共振能量转移技术观测MDCK上皮微组织的形成,检验不同的细胞力学信号和胞外基质环境对微组织发生的影响以及相应的ERK活性变化。结果与结论:①抑制ERK信号通路对MDCK上皮微组织的生长有抑制作用;②抑制细胞收缩力信号如ROCK通路和肌球蛋白MyosinⅡ活性,ERK活性降低,微组织变小;③选择性地抑制质膜和内质网上的钙离子通道导致ERK活性和微组织生长受到抑制;④通过抑制细胞膜上力学敏感受体Piezo离子通道或整合素信号,分别导致微组织变小或不能有效生成组织形态;⑤不同的细胞外基质组分影响微组织的形态发生,Matrigel胶中添加Ⅰ型胶原使微组织从微球形向长体形转变;⑥该研究结果初步显示细胞和细胞外基质环境中的机械力学信号在MDCK上皮微组织发生中具有重要作用,并提供了部分的分子机制理解。BACKGROUND:The development of tissues and organs in the body is a precise and autonomously regulated process,and the function of biomechanical factors at this macroscale is a basic scientific question worth exploring.OBJECTIVE:To investigate the roles of cell mechanics in morphogenesis of the lobular organoid of 3D Madin-Darby canine kidney(MDCK).METHODS:The formation of MDCK lobular organoid was visualized by fluorescence resonance energy transfer technology,and the influence of different cellular mechanical signals and extracellular matrix environment on lobular organoid formation and corresponding changes in extracellular regulated protein kinases(ERK)activity were examined.RESULTS AND CONCLUSION:(1)Inhibition of ERK signaling pathway can inhibit the growth of MDCK lobular organoid.(2)Inhibition of cell contractile force signals such as ROCK pathway and Myosin II activity,reduced ERK activity and lobular organoid size.(3)Selective inhibition of calcium channels in plasma membrane and endoplasmic reticulum led to reduced ERK activity and lobular organoid growth.(4)By inhibiting the mechanically-sensitive receptor Piezo ion channel or integrin signal on the cell membrane,the lobular organoid became smaller or MDCK cells could not generate tissue morphology.(5)Extracellular matrix compositions affected the morphogenesis of lobular organoid.The addition of type I collagen in Matrigel changed the lobular organoid to elongated shape.(6)The results of this study preliminarily show that mechanical signals in the cells and extracellular matrix environment play an important role in culturing MDCK lobular organoid,and provides certain molecular mechanisms.

关 键 词：ERK激酶 MDCK微组织 荧光共振能量转移 细胞力学 细胞外基质 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] Q28[生物学—细胞生物学]