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目的比较不同保存阶段红细胞及囊泡上相关蛋白组分的表达差异,积累红细胞老化、凋亡的相关信息。方法以10 d为1个周期,对保存红细胞制备红细胞血影和红细胞囊泡,SDS-PAGE电泳和双向电泳分析血影和囊泡的电泳图谱格局差异,免疫印迹检测GPA、GPC、stomatin、CD47、flotillin-1、flotillin-2、annexinⅦ和caveolin-1在血影及红细胞上的表达量。结果红细胞保存d11、d21、d31和d41时的囊泡的总蛋白量分别为180、483、1 055和1 357μg;血影和囊泡在PAGE电泳上的格局存在明显差异;d1、d21血影及d41囊泡的双向电泳图上分别可检出305、144和293个点;免疫印迹检测显示,GPC含量和保存时间之间无明显规律,其他几种蛋白的含量变化具有类似的趋势,即随着保存时间的增加,相应蛋白的含量在红细胞上不断减少,在囊泡上不断增加。结论随着保存时间的延长,红细胞多种蛋白表达量下调,囊泡是这些蛋白离开红细胞的1个重要载体。老化红细胞的清除机理可能是涉及多种表达下调膜蛋白的复杂机制。
Objective To compare the expression of related proteins in erythrocytes and vesicles at different storage stages and accumulate the related information of erythrocyte aging and apoptosis. Methods The erythrocyte ghost and erythrocyte vesicles were preserved for 10 d. The electrophoretogram of ghost and vesicle were analyzed by SDS-PAGE and two-dimensional gel electrophoresis. The expression of GPA, GPC, stomatin, CD47 , Flotillin-1, flotillin-2, annexinⅦ and caveolin-1 on the blood shadow and erythrocyte expression. Results The total amount of vesicles in the erythrocytes stored at d11, d21, d31 and d41 was 180,483,1 055 and 1 357μg, respectively. There was a significant difference in the pattern of blood vessels and vesicles on PAGE electrophoresis. The values of d1 and d21 And d41 vesicles were detected on the two-dimensional electrophoresis were 305,144 and 293 spots; Western blot analysis showed no significant difference between the GPC content and storage time, the content of several other proteins showed a similar trend, namely With the increase of storage time, the content of the corresponding protein decreased continuously on the red blood cells and increased on the vesicles. Conclusions With the prolongation of storage time, the expression of multiple proteins in erythrocytes is down-regulated. Vesicles are an important carrier of these proteins leaving erythrocytes. The mechanism of the clearance of aged erythrocytes may involve a number of complex mechanisms involved in the down-regulation of membrane proteins.