全血pH环境对红细胞膜流动性和膜蛋白的影响.docVIP

? 论 著 ? 5 全血pH环境对红细胞膜流动性和膜蛋白的影响 高美霞 吴敏慧 纪云鹏 蔡 莉 王泰瑞 杨 莹 梁文飚* （江苏省血液中心，江苏 南京 210042） 【摘要】目的 研究全血红细胞胞外pH环境对红细胞膜流动性和膜蛋白的影响。方法 将全血红细胞悬浮在不同pH值的ACD-B保养液中，分别在4、8和24h，利用荧光探针1，6-二苯基-1，3，5-己三烯（DPH）和SDS电泳检测红细胞膜流动性及红细胞膜蛋白；利用透射电镜观察红细胞形态；利用比色法检测红细胞上清中游离血红蛋白（FHb）。结果 除了环境pH在6.8的实验组外，其余各组的红细胞的偏振度、微黏度、Band3蛋白、红细胞形态和FHb与正常对照组存在差异性显著。结论 全血pH环境可影响红细胞膜流动性和膜蛋白的表达，可对红细胞的功能产生影响。【关键词】pH；红细胞；膜流动性；膜蛋白 中图分类号：R331 文献标识码：B 文章编号：1671-8194（2011）13-0005-03 Effects of Suspension pH and Storage Time on the Erythrocyte Membrane Fluidity and Membrane Protein GAO Mei-xia, WU Min-hui, JI Yun-peng, CAI Li, WANG Tai-rui, YANG Ying, LIANG Wen-biao (Jiangsu Province Blood Centre, Nanjing 210042, China) [Abstract] Objective To study the effects of suspension pH and storage time of whole blood on the erythrocyte membrane ? uidity and membrane protein. Methods Whole blood erythrocytes were suspended in different pH acid citrate dextrose B solution and their membrane fluidity, membrane protein, morphology of erythrocyte membrane and concentration of free hemoglobin(FHb) were assessed at the time of 4h, 8h and 24h by means of 1, 6-diphenyl-1, 3, 5-hexatriene(DPH) ? uorescence probe, SDSelectrophoresis, transmissionel ectron microscope and spectroscopy respectively. Results Except for pH 6.8 experimental group, there is a signi? cant difference in ? uorescence polarization(P), mean microviscosity (η), Band3 protein and FHb on comparaison with the control group. Conclusion Suspension pH and storage time of whole blood may in? uence the erythrocyte membrane ? uidity, membrane protein expression and the morphology of erythrocyte membrane, may produce affects on the red blood cell function.[Key words] pH; Erythrocyte; Membrane ? uidity; Membrane protein 红细胞悬液由全血制备而来，是目前临床输血中使用最多的红细胞类血液制剂。本研究探讨了全血红细胞用于MAP红悬液制备前，其细胞外pH和作用时间对红细胞膜流动性、膜蛋白和形态的影响，为探讨某些特殊情况的血液（如不足量血）制备红细胞悬液的应用潜能提供依据。1 材料与方法1.1 仪器及试剂 HITACHI KY2000型半自动生化分析仪、JEM-1010透射电镜、岛津RF-540型荧光分光光度计、NanoDrop ND-1000微量紫外可见分光光度计、Bio-Rad电泳仪及Quantity One分析软件；ACD-B全血保养液为山东威高