细胞变形性的能量学特征研究：基于原子力显微镜加载-卸载函数曲线分析

The Project proposes to study on the energy characteristics of cell deformation, based on analysis of loading-unloading curve, setting up deformability analysis system of three kinds of cells (RBC, HUVEC and hepatic cell) in the atomic force microscopy laboratory. Cell passive and active deformation was linkage analyzed; Under certain loading - unloading conditions, the theoretical relationship between the loading-unloading function curve will be researched, determine how to reflect and characterize the cell deformation by using loading-unloading function curve: (1) Cell deformability is the general characteristics in all biological cells; it is characterized by cell passive deformation and active deformation process together constitute; (2) cell deformation belong to energy index (Capacity), composed of integral difference of cell mechanical loading-unloading curve.The aim of investigating the energy characteristic of cell deformation is to make clear about important physical significance of cell mechanics parameters and cell deformability. Through this project research, to explain the mechanical behavior of a variety of cells, such as RBC deformation, HUVEC aligned along blood downstream and HSC fibrosis), to provide data and analysis system for new foundation. Through this project research, to explain the mechanical behavior of a variety of cells (RBC, HUVEC downstream of the deformation field aligned and HSC fibrosis) research to provide data and analysis system of new foundation.

以往细胞变形性指细胞的被动变形，忽略了主动变形。主动变形也是构成细胞变形性的重要内容，反映细胞伪足延伸和细胞迁移游走。本课题基于加载-卸载函数实验曲线分析，拟搭建维持二氧化碳孵育可用于原子力显微镜分析的实验平台。建立RBC、HUVEC和肝星状细胞HSC变形性分析系统。进行细胞被动-主动变形分析；研究加-卸载函数曲线理论关系，确定以加-卸载函数曲线表征细胞变形性：（1）细胞变形性由被动和主动变形共同构成；（2）细胞变形性具有能量特征，由细胞力学加载函数曲线与卸载函数曲线积分环面积所构成。研究细胞可变形性的能量特征及其物理意义。（3）用已建立的微管吸吮细胞系统，测定细胞力学参数进行对照分析；（4）红细胞激光衍射图像分析和血液流变学分析，研究单个细胞力学分析与宏观流变学指标的相关性；用已建立的荧光标记细胞结构蛋白原位分析技术，研究细胞变形的结构蛋白分子机制。

细胞变形性作为生物细胞的固有属性，在自然界中具有普遍性，它决定了细胞的形态和功能，在其适应特殊的生理环境中具有重要的科学意义。而我们对细胞变形性的认识一般是指细胞的被动变形，忽略了主动变形。其中，主动变形也是构成细胞变形性的重要内容，反映细胞迁移游走和伪足延伸等重要的生理现象。本课题中我们基于对原子力测得的力-距离曲线进行加载-卸载函数实验曲线分析，建立以红细胞为主的无核细胞以及以肝癌细胞为主的有核细胞的变形性分析系统。并构建了红细胞体内衰老模型和红细胞体外衰老模型，通过对其力学性质、膜骨架蛋白以及细胞内ATP水平的测定，探讨了红细胞衰老过程中变形性参数的能量学特征。通过对细胞被动变形-主动变形的联动研究分析，明确了细胞变形性的物理意义，理清了以往关于细胞力学特性的初步认识，也有助于深入认识细胞形态及其变化的生理意义，同时也为深入研究细胞生物力学特性的生理病理意义提供了基础。