QCM实时监测两种细胞骨架药物对烟草BY-2细胞粘弹性的影响  

Real-time Monitoring of Effects of Two Cytoskeleton Drugs on the Viscoelasticity of Tobacco(Nicotiana tabacum)BY-2 Cells by QCM

作　　者：兰雅琴 周铁安[1,2] 陈宗星 苏招红[2,3] 潘炜松 刘艺璇 LAN Ya-Qin;ZHOU Tie-An;CHEN Zong-Xing;SU Zhao-Hong;PAN Wei-Song;LIU Yi-Xuan(College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha 410128,China;Hunan Engineering Research Center for Cell Mechanics and Functional Analysis,Changsha 410128,China;College of Chemistry and Materials Science,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学生物科学技术学院,长沙410128 [2]湖南省细胞力学与功能分析工程技术研究中心,长沙410128 [3]湖南农业大学化学与材料科学学院,长沙410128

出　　处：《农业生物技术学报》2022年第2期305-315,共11页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31870933);湖南省自然科学基金(2018JJ2164)。

摘　　要：植物细胞骨架在植物生长发育与不同环境胁迫下将进行动态重塑,从而导致细胞骨架力学性能的变化。本研究旨在使用细胞骨架药物改变细胞骨架的稳定性,进而从细胞骨架结构层次研究烟草(Nicotiana tabacum)BY-2细胞的粘弹性。采用石英晶体微天平(quartz crystal microbalance,QCM)技术实时监测了BY-2细胞在2种细胞骨架药物作用下的粘弹性响应。结果表明,用紫杉醇处理细胞,在低浓度时(10,15,17μmol/L),细胞的细胞粘弹性指数(cell viscoelastic index,CVI)先减小再增大,细胞先变软再变硬;在20和25μmol/L紫杉醇作用下,细胞的CVI由负值至最终都接近0,细胞变硬且硬度比加药前大。用细胞松弛素D处理细胞时,在低浓度时(50,100,150 ng/mL),CVI由加药前的负值增大至远大于0后回落,细胞先变软再变硬;在高浓度(200,250 ng/mL)时,CVI由负值增大至稍小于0,细胞先快速变硬后变软,但硬度比加药前大。除药物对微管与微丝的直接作用外,用细胞骨架张力整合模型以及赫氏斑通过回缩引起质膜张力增加解释了微管与微丝在药物作用下的细胞软硬度变化。此外,使用荧光显微镜观测了200 ng/mL细胞松弛素D作用下烟草细胞微丝结构的变化,观测结果与QCM实时监测得到的结果一致。本研究所建立的方法为通过细胞粘弹性的测定研究植物细胞层次结构提供了新的途径,为进一步研究植物细胞在细胞骨架药物作用下的力学性能提供了基础。Plant cytoskeleton undergoes dynamic remodeling under plant growth and development and different environmental stresses,resulting in changes in the mechanical properties of the cytoskeleton.Cytoskeleton drugs can change the stability of the cytoskeleton,so cytoskeleton drugs can be used to study the viscoelasticity of tobacco(Nicotiana tabacum)BY-2 cells from the level of cellular structure.In this paper,quartz crystal microbalance(QCM)was used to monitor the viscoelastic response of BY-2 cells under the treatments of two cytoskeleton drugs in real time.The results showed that when the cells were treated with low concentrations of paclitaxel(10,15,17μmol/L),the cell viscoelastic index(CVI)of the cells decreased and then increased,and the cells first became softer and then harder.This softening was almost instantaneous,reflecting the rapid force signal participation process.Under the actions of 20 and 25μmol/L paclitaxel,the CVI of the cells went from negative values to finally close to zero,and the cells became harder and the hardness was greater than that before the drug was added.Under the treatments of lower concentrations of drug cytochalasin D,at low concentrations(50,100,150 ng/mL),the CVI of the cells increased from the negative values before adding the drug to greater than zero then dropped,and the cells first became softer and then became harder,the rapid change of CVI’s numerical sign reflected the rapid and unstable change of the cells’generated force;and at higher concentrations(200,250 ng/mL),the CVI of the cells went from negative values to finally slightly greater than zero,and the cells quickly became harder and then became softer,but the hardness was larger than that before adding the drugs.Cellular tensegrity model and the increased plasma membrane tension caused by retraction of the Hechtian strands were used to explain the changes in cells’softness or stiffness under the actions of the two cytoskeleton drugs.In addition,microscopy technology was used to observe the changes in the micro

关 键 词：石英晶体微天平(QCM) 烟草BY-2细胞 粘弹性 紫杉醇 细胞松弛素D 

分 类 号：S313[农业科学—作物栽培与耕作技术]