剪切应力对晚期内皮祖细胞细胞骨架蛋白表达及分布的影响  

Effects of shear stress on the expression and location of cytoskeletal proteins in late endothelial progenitor cells

作　　者：张怡 王信[2] 成敏 黄厚今 Zhang Yi;Wang Xin;Cheng Min;Huang Houjin(Department of Preventive Medicine,School of Public Health,Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Orthopaedic Surgery,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563099,China;Medicine Research Center,Weifang Medical University,Weifang Shandong 261053,China)

机构地区：[1]遵义医学院公共卫生学院卫生毒理学教研室,贵州遵义563099 [2]遵义医学院附属医院骨科,贵州遵义563099 [3]潍坊医学院医学研究实验中心,山东潍坊261053

出　　处：《遵义医学院学报》2018年第4期413-417,共5页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:30900290;31270993)

摘　　要：目的探讨剪切应力对晚期内皮祖细胞(endothelial progenitor cells,EPCs)细胞骨架蛋白局部粘着斑激酶(focal adhesion kinase,FAK)、踝蛋白(talin)和桩蛋白(paxillin)表达及分布的影响。方法梯度密度离心法获取大鼠骨髓单核细胞,体外培养EPCs并鉴定。以第三代即晚期EPCs作为靶细胞,给予12 dyne/cm^2的剪切应力干预3 h,以静止组作为空白对照。分别采用Western Blot和细胞免疫荧光观察3种细胞骨架蛋白FAK、talin和paxillin表达量和分布情况。结果与静止组相比,剪切应力作用EPCs 3 h后:FAK蛋白的表达量显著增加,变化为静止组的2.06±0.01倍,差异有统计学意义(P<0.01),表达位置沿剪切应力方向聚集成束并延伸至细胞边缘;而talin和paxillin蛋白的表达量则均有所下降(P<0.01)。结论剪切应力能影响晚期EPCs上细胞骨架蛋白FAK、talin和paxillin的表达及分布。Objective To investigate the effects of shear stress on the expression and location of cytoskeletal proteins,such as Focal adhesion kinase （FAK） ,talin,and paxillin in late endothelial progenitor cells. Methods Mononuclear cells were isolated from rat bone marrow by density gradient centrifugation and cultured with specific culture medium （M199 medium contained with VEGF、bFGF and 15% bovine serum）. The EPCs were identified by Dil-ac-LDL and FITC-UEA-1 double staining. Micro-fluidic platform was used to simulate the shear stress generated by blood flow and tissue fluid flow. EPCs were either exposed to the shear stress （12 dyne/cm^2）for 3 h or kept in static condition （the control group）. The protein levels of FAK,talin,and paxillin were detected by Western Blot （WB） and the distributions of these proteins were visualized using Immunofluorescence （IF）.Results Compared with the control group,WB results showed that the protein levels of talin and paxillin were significantly decreased in the shear stress group but the expression of FAK protein was increased （P〈0. 01）. Similarly,the fluorescence intensities of FAK were obviously enhanced in EPCs exposed to shear stress. However,substantial decreases in the fluorescence intensities of talin and paxillin were detected under shear stress conditions.Moreover,it has been found that shear stress induced the peripheral redistribution of FAK protein in EPCs.Conclusion Shear stress can induce the expression and redistribution of cytoskeletal proteins in late EPCs.

关 键 词：晚期内皮祖细胞 剪切应力 局部粘着斑激酶 踝蛋白 桩蛋白 

分 类 号：R318[医药卫生—生物医学工程]