基底刚度对新生大鼠心脏成纤维细胞增殖和粘附的影响  

Effects of substrate rigidity on proliferation and adhesion in neonatal rat cardiac fibroblasts

作　　者：高炜[1] 秦雷[1] 熊春阳[2] 方竞[2] 张幼怡[1] 

机构地区：[1]北京大学第三医院血管医学研究所分子心血管学教育部重点实验室,北京100191 [2]北京大学前沿交叉学科研究院,北京100871

出　　处：《中国分子心脏病学杂志》2010年第2期75-80,共6页Molecular Cardiology of China

基　　金：国家重点基础研究发展规划项目计划(2007CB935602);国家自然科学基金国际合作与交流项目(30910103902)

摘　　要：目的探讨细胞培养基底刚度对心肌成纤维细胞增殖和粘附的影响。方法制备不同配比聚丙烯酰胺凝胶模拟不同基底刚度,采用细胞计数和MTT法检测细胞增殖,免疫荧光成像和免疫印迹方法检测细胞粘着斑分布及其相关蛋白表达。结果采用自重弯曲法测得0.03%、0.13%和0.26%聚丙烯酰胺凝胶的弹性模量为7～33Kpa。在此刚度范围内,随着基底刚度增加心脏成纤维细胞数量显著增加,粘着斑面积占细胞总面积的百分比增加,粘着斑纽蛋白的荧光强度增加,但纽蛋白和桩蛋白的表达量不变。结论基底刚度增加能促进新生大鼠心脏成纤维细胞的增殖和改变粘着斑蛋白的分布。Objective To observe the effects of substrate rigidity on proliferation and adhesion in neonatal rat cardiac fibroblasts. Methods Polyaerylamide gels with different ratio （Bis：Acr） were prepared to achieve different stiffness substrates for cell culture. The cell proliferation was determined by using cell counting and MTT assay. The distribution and expression of focal adhesion proteins were detected by immunofluorescence confocal image and Western Blotting. Results Young＇ s modulus of the substrate gel meas ured by the methods of micro-ball indentation, gravity stretching and special apparatus. Young＇s modulus E value of the gel substrate ranged from 7 KPa to 33KPa. The variation of cell number, percentages of focal adhesion area in ceil area, and fluorescence intensity of vinculin increases, but not expression of vinculin and paxillin , along with substrate stiffness increase. Conclusion Substrate stiffness increasing may contribute to cell proliferation and focal adhesion formation in neonatal rat cardiac fibroblasts.

关 键 词：刚度 增殖 粘着斑 心脏成纤维细胞 

分 类 号：R722.1[医药卫生—儿科]