流体粘度对大鼠脑微血管内皮细胞微区力学特性的影响
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摘要
失血性休克是临床常见的休克类型。液体复苏是治疗失血性休克的重要措施,旨在增加有效循环血量、改善微循环,减轻组织和器官的损伤。液体复苏所采用的血浆扩容剂不仅机械性的扩充血容量,而且影响血浆粘度和其他流变学特性,进一步影响组织的血供、氧供和微循环,关系到组织器官功能的恢复。传统的观点曾认为,血液粘度升高不利于血流灌注,所以应该选择低粘度的血浆扩容剂进行复苏。而上个世纪90年代开始逐渐形成的观点认为,在重度失血性休克或极度血液稀释等情况下,应该选用高粘度的血浆扩容剂,后者能够维持微血管血流,改善血流动力学指标,提高失血性休克的复苏效果。曾有研究者根据动物实验的结果推测,高粘度血浆扩容剂可能通过血浆粘度上升引起微血管壁剪应力升高,微血管内皮细胞舒张因子释放增加,改善了微循环的血流灌注。但动物整体水平研究限制了机制研究的深入,高粘度复苏的观点尚未得到普遍接受,重度失血性休克早期救治中适宜的血浆粘度范围仍存在争议。所以,流体粘度对微血管内皮细胞功能的影响及其机制有待深入探究。
微血管内皮细胞处在复杂的力学环境中,承受多种力学刺激。细胞刚度、弹性和粘滞性等力学特性的变化既是力学刺激的结果,又是细胞功能维持的基础,对微循环血流灌注具有重要的意义。原子力显微镜(atomic force microscopy, AFM)能够在接近生理的条件下获得高分辨的三维形貌像,同时对细胞微区力学特性进行定量描述。本研究利用体外层流剪切体系和AFM微区力学特性的探测功能,目的在于深入探讨剪切速率恒定的情况下,不同流体粘度对微血管内皮细胞微区力学特性的影响,明确细胞骨架、NO和ET-1等在细胞力学特性变化中的可能作用。以期从生物力学角度揭示流体粘度对微血管内皮细胞的调控机制,为重度失血性休克早期紧急救治提供更加合理的理念,为血液和血浆代用品的研制提供必要的参考。
不同流体粘度大鼠脑微血管内皮细胞剪切体系的建立探讨流动剪切环境下粘度对血管内皮细胞力学特性的影响,需要首先建立体外模拟血流对内皮细胞施加剪应力的研究体系。本研究依据流体剪应力场分布的要求,设计加工了平行板流动小室,在使用方便性和性能两方面达到了国外商品化同类产品的水平。以平行板流动小室为核心,建立了封闭、稳定及剪切速率可调的层流剪切体系。进行了大鼠脑微血管内皮细胞(rat cerebral microvessel endothelial cell,rCMECs)的原代培养,经内皮细胞的特征性鉴定,纯度能够满足进一步实验的需要,且经40 dyn/cm2剪应力作用2h不发生脱落。海藻酸钠溶液表现近似牛顿流体的特性。与右旋糖酐70相比,海藻酸钠在浓度和胶体渗透压很低的情况下能够产生很高的粘度,对rCMECs的生长没有明显影响,适合做“粘度调节剂”。所建立的不同流体粘度微血管内皮细胞剪切体系能够满足进一步研究的需要。
AFM定量研究rCMECs微区力学特性的参数确定Force Volume(群体力谱)是AFM研究样品微区力学特性的常用工作模式,在针尖以接触模式扫描样品表面的同时采集每个象素点的力曲线。杨氏模量是目前定量描述细胞微区力学特性的常用参数,本研究确定了基于Hertz模型的、适宜本研究体系的rCMECs微区杨氏模量计算参数。另外,首次将基于力曲线的形变消耗功(Deformation Consumed Work)引入rCMECs微区力学特性的分析,其数值反映细胞形变与理想弹性体形变消耗功的差异,与细胞的粘弹性直接相关。与Hertz模型计算杨氏模量相比,形变消耗功适合于描述细胞非线性弹性形变,基本不受基底的影响,且数值不随压入深度变化而改变,作为新的定量参数,有可能成为Hertz模型计算杨氏模量的补充。血管内皮细胞的形变消耗功能够在一定程度上反映心脏做功在血管壁的能量消耗。
流体粘度对rCMECs形态和微区力学特性的影响在剪切速率为800s-1,灌流液的粘度分别为0.93,2.08和4.76 mPa s(剪应力分别为7.4,16.6和38.1 dyn/cm2)作用2h后,rCMECs方向角分布的变化表明细胞有沿剪应力作用方向取向的趋势,高粘度灌流液作用引起一些细胞伸长变细。随流体粘度的升高,剪应力增大,细胞厚度降低、变平坦,rCMECs微区力学特性也发生明显变化。表现为刚度上升,形变消耗功和表面非特异性粘滞力降低。但力学特性的变化有一定限度,当流体粘度和剪应力上升到一定程度,力学特性不会再继续发生变化。rCMECs微区刚度上升是细胞骨架重组加剧的结果,形变消耗功和表面非特异粘滞力的降低,提示心脏做功消耗在血管内皮细胞上的能量以及白细胞与血管内皮细胞之间非特异性黏附的发生几率可能随血液粘度升高而降低。以上结果说明,适当增加血液粘度能够通过调节微血管内皮细胞的力学特性而降低血流在微循环的能量消耗,减少白细胞黏附,改善微循环血流灌注。但体内血液粘度过高不仅不能改善内皮细胞的力学特性,反而可能导致血流速度下降、剪应力降低,引起内皮细胞损伤。
流体粘度对rCMECs细胞骨架肌动蛋白重组和舒缩因子生成的影响在剪切速率一定的情况下,随流体粘度增大、剪应力升高,rCMECs内细胞骨架F-actin分布发生变化,核区应力纤维束增加。β-actin mRNA水平急剧增加反映了rCMECs对不同强度剪应力的适应,表现了肌动蛋白更新、重组速率加快以及细胞向基底的黏附力增强。ET-1在mRNA水平的表达随剪应力强度的增大而升高,当达到一定的阈值后,会随剪应力的继续增大而降低。灌流液中NO的增加与F-actin的重新分布相伴。以上结果说明,细胞骨架肌动蛋白是流体粘度引起rCMECs力学特性变化的物质基础,NO的分泌与细胞骨架的重组密切相关。
综上所述,本研究建立了体外不同流体粘度微血管内皮细胞的层流剪切体系,确定了基于AFM力曲线的、适用于rCMECs的微区力学特性计算参数。观察到在相同剪切速率下,在一定范围内,随流体粘度的增加,rCMECs的三维形态和微区力学特性发生了明显的变化。说明在重度失血性休克的早期救治中,一定程度上提高血液或血浆粘度能够通过提高剪应力调节微血管内皮细胞的力学特性,降低心脏做功的能量消耗,引起微血管扩张,改善微循环。
Hemorrhagic shock remains a major cause of death and disability in battlefield injuries, as well as in civilian trauma. Fluid resuscitation is an essential component of therapy for hemorrhagic shock. The aim of fluid resuscitation is to expend effective blood volume, increase perfusion in microcirculation, and to alleviate injuries in tissue and organ. Plasma expanders used in fluid resuscitation can not only expend the blood volume, but also influence plasma viscosity and other rheological behaviors, which are important determinants in tissue perfusion, oxygen supply, microcirculation and improvement in function of tissue and organs. It was generally perceived that lowered blood viscosity in hemodilution and hemorrhagic shock could improve the tissue perfusion due to reduced flow resistance. But the studies began in 90th in the last century showed that plasma expander with high viscosity, which could increase perfusion in microcirculation and improve hemodynamic parameters, is beneficial in extreme hemodilution and severe hemorrhagic shock. Plasma expander with high viscosity is known to elvate plasma viscosity and wall shear stress in microvascular, and to improve function of endothelial cell in microvascular. But because of the limitation in animal study, the effects of hydro- viscosity on microvessel endothelial cell and the mechanisms are still to be investigated.
Microvessel endothelial cells locate in circumstances with complex mechanical forces. Changes in mechanical properties, such as stiffness, elasticity, and adherent property are outcomes of mechanical stimulation, as well as maintenance of cellular function. Mechanical properties of microvessel endothelial cells are important in perfusion of microcirculation. Atomic force microscopy (AFM) is a novel method for high-resolution three-dimensional topography in living cell, and can detect local mechanical properties of cells. The present study will use in vitro perfusing system of luminar flow and function of detecting local mechanical properties by AFM. The aim of the study is to observe the influence of hydro- viscosity on the mechanics of microvessel endothelial cell, and to investigate the roles of cytoskeleton, NO (nitric oxide) and ET-1 (Endothelin-1). The regulatory mechanisms of hydro-viscosity on the mechanics of microvessel endothelial cell will give more insight in the treatment of hemorrhagic shock and the research in blood and plasma substitutes.
Establishment of system of rat cerebral microvessel endothelial cells perfused with medium of different hydro-viscosities. To investigate he influence of hydro- viscosity on the mechanics of microvessel endothelial cell, it is necessary to establish an in vitro system to simulate shear stress of blood flow. We developed a parallel plate flow chamber according to the fundamental of distribution of shear stress, which showed equally in ease of use and performance compared with overseas commercial chambers. Based on the parallel plate flow chamber, in vitro perfusing system of luminar flow was developed. Rat cerebral microvessel endothelial cells (rCMECs) were isolated and cultured in vitro. The purity of the cells meets the demand in further study and can keep stable in the shear stress. Sodium alginate solutions show Newton property. Compared with Dextran 70, sodium alginate of a comparatively low concentration can result in high viscosity with a low colloid oncotic pressure and a low concentration. Sodium alginate solutions do not influence the growth of rCMECs. Thus, sodium alginate can act as‘viscosity modifier’. The system of microvessel endothelial cells perfused with medium of different hydro-viscosities meet the demand in further study.
Determination of parameters for local mechanical properties of rCMECs in quantitative analysis by AFM. Force volume is the commonly used working pattern for detection of local mechanical properties by AFM. The Force-Curve can be collected during the scanning of surface of the cells by contact mode. Young’s modular is the commonly used parameter in quantitative analysis of local mechanical properties. Method to evaluate Young’s modular suitable for rCMECs in the present was developed based on Hertz model. Meanwhile,‘Deformation Consumed Work’calculated based on the Force-Curve was advanced for the first time in this study for analyzing the local mechanical properties. Deformation Consumed Work, which shows viscoelasticity of cells, describes the differences in Consumed Work between deformation of the cell and elastic homogeneous body. Deformation Consumed Work is suitable in quantitative analysis of nonlinear deformation of cells. Furthermore, it can eliminate the influence of hard base and show no changes with different indetations. As a novel parameter in quantitative analysis of local mechanical properties, Deformation Consumed Work may be an alternative or compensatory of Young’s modular calculated by Hertz model. Deformation Consumed Work of endothelial cell can reflect the energy dissipation of blood flow in wall of vascular generated by heart.
Effect of hydro-viscosity on morphology and local mechanical properties of rCMECs. Under the shear rate of 800s-1, hydro-viscosity of 0.93, 2.08 and 4.76 mPa s, shear stress of 7.4, 16.6 and 38.1 dyn/cm2 for 2h, the changes of distribution of angle of rCMECs reflect orientation caused by the shear stress. Higher viscosity causes some cells to become thinner and longer. As the hydro-viscosity and shear stress become higher, the height and the roughness of the cell decrease. Meanwhile, the stiffness elevates, but Deformation Consumed Work and non-specific adherent force decrease. The changes of local mechanical properties have threshold. When the hydro-viscosity reaches a certain level, local mechanical properties do not show more changes. The elevate stiffness is the result of reorganization of cytoskeleton. Decrease in Deformation Consumed Work reflects reduction in energy dissipation of blood flow in wall of vascular generated by heart. Decrease in non-specific adherent force reflects lower frequency of adhesion of white blood cell to endothelial cell. These results indicated that appropriately elevated hydro-viscosity can modify local mechanical properties of rCMECs, and can reduce energy dissipation in microcirculation, restrain the adhesion of white blood cell to endothelial cell. These mechanisms are beneficial in perfusion in microcirculation. But if the blood viscosity becomes too high in vivo, the local mechanical properties of endothelial cells cannot improve.
Effect of hydro-viscosity on cytosleletal actin reorganization and production of NO and ET-1 in rCMECs. At a constant shear rate, higher viscosity causes changes in distribution of cytoskeleton F-actin, and stress fibers become dense in the central region of rCMECs. The dramatic increase ofβ-actin mRNA reflects response of rCMECs to shear stress, which includes a greater turnover of actin, and stronger adherent force to basement. ET-1 mRNA increases when hydro-viscosity and shear stress become higher. But if hydro-viscosity and shear stress reach a certain threshold, the ET-1 mRNA decreases. The increases of NO production show concomitance with the F-actin reorganization. These results indicated that cytosleletal actin is the base of changes in local mechanical properties, and NO production shows relationship with cytoskeletal reorganization.
Taken together, system of microvessel endothelial cells perfused with medium of different hydro-viscosities was established. Parameters for local mechanical properties in quantitative analysis by AFM were determined. Three-dimensional topography and local mechanical properties of rCMECs changed significantly when hydro-viscosity became higher and the shear rate kept constant. The regulatory mechanisms of hydro-viscosity of certain degree on the mechanics of microvessel endothelial cell will be beneficial in the treatment of severe hemorrhagic shock and the viscosity of blood and plasma substitutes should be taken into consideration.
微血管内皮细胞处在复杂的力学环境中,承受多种力学刺激。细胞刚度、弹性和粘滞性等力学特性的变化既是力学刺激的结果,又是细胞功能维持的基础,对微循环血流灌注具有重要的意义。原子力显微镜(atomic force microscopy, AFM)能够在接近生理的条件下获得高分辨的三维形貌像,同时对细胞微区力学特性进行定量描述。本研究利用体外层流剪切体系和AFM微区力学特性的探测功能,目的在于深入探讨剪切速率恒定的情况下,不同流体粘度对微血管内皮细胞微区力学特性的影响,明确细胞骨架、NO和ET-1等在细胞力学特性变化中的可能作用。以期从生物力学角度揭示流体粘度对微血管内皮细胞的调控机制,为重度失血性休克早期紧急救治提供更加合理的理念,为血液和血浆代用品的研制提供必要的参考。
不同流体粘度大鼠脑微血管内皮细胞剪切体系的建立探讨流动剪切环境下粘度对血管内皮细胞力学特性的影响,需要首先建立体外模拟血流对内皮细胞施加剪应力的研究体系。本研究依据流体剪应力场分布的要求,设计加工了平行板流动小室,在使用方便性和性能两方面达到了国外商品化同类产品的水平。以平行板流动小室为核心,建立了封闭、稳定及剪切速率可调的层流剪切体系。进行了大鼠脑微血管内皮细胞(rat cerebral microvessel endothelial cell,rCMECs)的原代培养,经内皮细胞的特征性鉴定,纯度能够满足进一步实验的需要,且经40 dyn/cm2剪应力作用2h不发生脱落。海藻酸钠溶液表现近似牛顿流体的特性。与右旋糖酐70相比,海藻酸钠在浓度和胶体渗透压很低的情况下能够产生很高的粘度,对rCMECs的生长没有明显影响,适合做“粘度调节剂”。所建立的不同流体粘度微血管内皮细胞剪切体系能够满足进一步研究的需要。
AFM定量研究rCMECs微区力学特性的参数确定Force Volume(群体力谱)是AFM研究样品微区力学特性的常用工作模式,在针尖以接触模式扫描样品表面的同时采集每个象素点的力曲线。杨氏模量是目前定量描述细胞微区力学特性的常用参数,本研究确定了基于Hertz模型的、适宜本研究体系的rCMECs微区杨氏模量计算参数。另外,首次将基于力曲线的形变消耗功(Deformation Consumed Work)引入rCMECs微区力学特性的分析,其数值反映细胞形变与理想弹性体形变消耗功的差异,与细胞的粘弹性直接相关。与Hertz模型计算杨氏模量相比,形变消耗功适合于描述细胞非线性弹性形变,基本不受基底的影响,且数值不随压入深度变化而改变,作为新的定量参数,有可能成为Hertz模型计算杨氏模量的补充。血管内皮细胞的形变消耗功能够在一定程度上反映心脏做功在血管壁的能量消耗。
流体粘度对rCMECs形态和微区力学特性的影响在剪切速率为800s-1,灌流液的粘度分别为0.93,2.08和4.76 mPa s(剪应力分别为7.4,16.6和38.1 dyn/cm2)作用2h后,rCMECs方向角分布的变化表明细胞有沿剪应力作用方向取向的趋势,高粘度灌流液作用引起一些细胞伸长变细。随流体粘度的升高,剪应力增大,细胞厚度降低、变平坦,rCMECs微区力学特性也发生明显变化。表现为刚度上升,形变消耗功和表面非特异性粘滞力降低。但力学特性的变化有一定限度,当流体粘度和剪应力上升到一定程度,力学特性不会再继续发生变化。rCMECs微区刚度上升是细胞骨架重组加剧的结果,形变消耗功和表面非特异粘滞力的降低,提示心脏做功消耗在血管内皮细胞上的能量以及白细胞与血管内皮细胞之间非特异性黏附的发生几率可能随血液粘度升高而降低。以上结果说明,适当增加血液粘度能够通过调节微血管内皮细胞的力学特性而降低血流在微循环的能量消耗,减少白细胞黏附,改善微循环血流灌注。但体内血液粘度过高不仅不能改善内皮细胞的力学特性,反而可能导致血流速度下降、剪应力降低,引起内皮细胞损伤。
流体粘度对rCMECs细胞骨架肌动蛋白重组和舒缩因子生成的影响在剪切速率一定的情况下,随流体粘度增大、剪应力升高,rCMECs内细胞骨架F-actin分布发生变化,核区应力纤维束增加。β-actin mRNA水平急剧增加反映了rCMECs对不同强度剪应力的适应,表现了肌动蛋白更新、重组速率加快以及细胞向基底的黏附力增强。ET-1在mRNA水平的表达随剪应力强度的增大而升高,当达到一定的阈值后,会随剪应力的继续增大而降低。灌流液中NO的增加与F-actin的重新分布相伴。以上结果说明,细胞骨架肌动蛋白是流体粘度引起rCMECs力学特性变化的物质基础,NO的分泌与细胞骨架的重组密切相关。
综上所述,本研究建立了体外不同流体粘度微血管内皮细胞的层流剪切体系,确定了基于AFM力曲线的、适用于rCMECs的微区力学特性计算参数。观察到在相同剪切速率下,在一定范围内,随流体粘度的增加,rCMECs的三维形态和微区力学特性发生了明显的变化。说明在重度失血性休克的早期救治中,一定程度上提高血液或血浆粘度能够通过提高剪应力调节微血管内皮细胞的力学特性,降低心脏做功的能量消耗,引起微血管扩张,改善微循环。
Hemorrhagic shock remains a major cause of death and disability in battlefield injuries, as well as in civilian trauma. Fluid resuscitation is an essential component of therapy for hemorrhagic shock. The aim of fluid resuscitation is to expend effective blood volume, increase perfusion in microcirculation, and to alleviate injuries in tissue and organ. Plasma expanders used in fluid resuscitation can not only expend the blood volume, but also influence plasma viscosity and other rheological behaviors, which are important determinants in tissue perfusion, oxygen supply, microcirculation and improvement in function of tissue and organs. It was generally perceived that lowered blood viscosity in hemodilution and hemorrhagic shock could improve the tissue perfusion due to reduced flow resistance. But the studies began in 90th in the last century showed that plasma expander with high viscosity, which could increase perfusion in microcirculation and improve hemodynamic parameters, is beneficial in extreme hemodilution and severe hemorrhagic shock. Plasma expander with high viscosity is known to elvate plasma viscosity and wall shear stress in microvascular, and to improve function of endothelial cell in microvascular. But because of the limitation in animal study, the effects of hydro- viscosity on microvessel endothelial cell and the mechanisms are still to be investigated.
Microvessel endothelial cells locate in circumstances with complex mechanical forces. Changes in mechanical properties, such as stiffness, elasticity, and adherent property are outcomes of mechanical stimulation, as well as maintenance of cellular function. Mechanical properties of microvessel endothelial cells are important in perfusion of microcirculation. Atomic force microscopy (AFM) is a novel method for high-resolution three-dimensional topography in living cell, and can detect local mechanical properties of cells. The present study will use in vitro perfusing system of luminar flow and function of detecting local mechanical properties by AFM. The aim of the study is to observe the influence of hydro- viscosity on the mechanics of microvessel endothelial cell, and to investigate the roles of cytoskeleton, NO (nitric oxide) and ET-1 (Endothelin-1). The regulatory mechanisms of hydro-viscosity on the mechanics of microvessel endothelial cell will give more insight in the treatment of hemorrhagic shock and the research in blood and plasma substitutes.
Establishment of system of rat cerebral microvessel endothelial cells perfused with medium of different hydro-viscosities. To investigate he influence of hydro- viscosity on the mechanics of microvessel endothelial cell, it is necessary to establish an in vitro system to simulate shear stress of blood flow. We developed a parallel plate flow chamber according to the fundamental of distribution of shear stress, which showed equally in ease of use and performance compared with overseas commercial chambers. Based on the parallel plate flow chamber, in vitro perfusing system of luminar flow was developed. Rat cerebral microvessel endothelial cells (rCMECs) were isolated and cultured in vitro. The purity of the cells meets the demand in further study and can keep stable in the shear stress. Sodium alginate solutions show Newton property. Compared with Dextran 70, sodium alginate of a comparatively low concentration can result in high viscosity with a low colloid oncotic pressure and a low concentration. Sodium alginate solutions do not influence the growth of rCMECs. Thus, sodium alginate can act as‘viscosity modifier’. The system of microvessel endothelial cells perfused with medium of different hydro-viscosities meet the demand in further study.
Determination of parameters for local mechanical properties of rCMECs in quantitative analysis by AFM. Force volume is the commonly used working pattern for detection of local mechanical properties by AFM. The Force-Curve can be collected during the scanning of surface of the cells by contact mode. Young’s modular is the commonly used parameter in quantitative analysis of local mechanical properties. Method to evaluate Young’s modular suitable for rCMECs in the present was developed based on Hertz model. Meanwhile,‘Deformation Consumed Work’calculated based on the Force-Curve was advanced for the first time in this study for analyzing the local mechanical properties. Deformation Consumed Work, which shows viscoelasticity of cells, describes the differences in Consumed Work between deformation of the cell and elastic homogeneous body. Deformation Consumed Work is suitable in quantitative analysis of nonlinear deformation of cells. Furthermore, it can eliminate the influence of hard base and show no changes with different indetations. As a novel parameter in quantitative analysis of local mechanical properties, Deformation Consumed Work may be an alternative or compensatory of Young’s modular calculated by Hertz model. Deformation Consumed Work of endothelial cell can reflect the energy dissipation of blood flow in wall of vascular generated by heart.
Effect of hydro-viscosity on morphology and local mechanical properties of rCMECs. Under the shear rate of 800s-1, hydro-viscosity of 0.93, 2.08 and 4.76 mPa s, shear stress of 7.4, 16.6 and 38.1 dyn/cm2 for 2h, the changes of distribution of angle of rCMECs reflect orientation caused by the shear stress. Higher viscosity causes some cells to become thinner and longer. As the hydro-viscosity and shear stress become higher, the height and the roughness of the cell decrease. Meanwhile, the stiffness elevates, but Deformation Consumed Work and non-specific adherent force decrease. The changes of local mechanical properties have threshold. When the hydro-viscosity reaches a certain level, local mechanical properties do not show more changes. The elevate stiffness is the result of reorganization of cytoskeleton. Decrease in Deformation Consumed Work reflects reduction in energy dissipation of blood flow in wall of vascular generated by heart. Decrease in non-specific adherent force reflects lower frequency of adhesion of white blood cell to endothelial cell. These results indicated that appropriately elevated hydro-viscosity can modify local mechanical properties of rCMECs, and can reduce energy dissipation in microcirculation, restrain the adhesion of white blood cell to endothelial cell. These mechanisms are beneficial in perfusion in microcirculation. But if the blood viscosity becomes too high in vivo, the local mechanical properties of endothelial cells cannot improve.
Effect of hydro-viscosity on cytosleletal actin reorganization and production of NO and ET-1 in rCMECs. At a constant shear rate, higher viscosity causes changes in distribution of cytoskeleton F-actin, and stress fibers become dense in the central region of rCMECs. The dramatic increase ofβ-actin mRNA reflects response of rCMECs to shear stress, which includes a greater turnover of actin, and stronger adherent force to basement. ET-1 mRNA increases when hydro-viscosity and shear stress become higher. But if hydro-viscosity and shear stress reach a certain threshold, the ET-1 mRNA decreases. The increases of NO production show concomitance with the F-actin reorganization. These results indicated that cytosleletal actin is the base of changes in local mechanical properties, and NO production shows relationship with cytoskeletal reorganization.
Taken together, system of microvessel endothelial cells perfused with medium of different hydro-viscosities was established. Parameters for local mechanical properties in quantitative analysis by AFM were determined. Three-dimensional topography and local mechanical properties of rCMECs changed significantly when hydro-viscosity became higher and the shear rate kept constant. The regulatory mechanisms of hydro-viscosity of certain degree on the mechanics of microvessel endothelial cell will be beneficial in the treatment of severe hemorrhagic shock and the viscosity of blood and plasma substitutes should be taken into consideration.
引文
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