摘要
经糖基化修饰可使4℃保存的血小板在输入体内后延长其存活。本研究探讨尿苷二磷酸半乳糖(UDP-Gal)糖基化修饰对血小板形态、结构、功能以及其膜糖蛋白的影响。实验分为室温对照组、冷藏对照组和修饰组。用荧光标记的特异凝集素(FITC-RCAⅠ)检测膜糖蛋白糖残基变化,用扫描、透射电镜观察修饰后血小板形态和超微结构的变化,用比浊法测定血小板聚集率,用流式细胞术检测血小板膜蛋白CD42b、膜表面标志CD62P以及血小板凋亡标志annexin Ⅴ结合率。结果表明,UDP-Gal修饰组RCAⅠ结合率显著高于室温对照组和冷藏对照组(p<0.01);与新鲜血小板相比,UDP-Gal处理后的血小板超微结构无明显变化,而冷藏对照组则有伪足延伸等形态学改变;修饰后最大聚集率可达新鲜血小板的50%以上;血小板膜蛋白CD42b、膜表面标志CD62P及annexin Ⅴ结合率与室温对照组相比均无明显差异。结论:UDP-Gal可使β-半乳糖有效地结合于链聚糖末端,糖基化血小板仍然具有较强的聚集活性和相对完整的超微结构,功能基本正常。
The glycosylation of platelests may prolong their life-span when being transfused after preservation under 4℃, therefore this study was aimed to investigate the effect of glycosylation on morphology, ultmstructure, function and membrane glycoprotein of platelets. The experiments were divided into 3 groups： group preserved in room temperature （ RT group）, group preserved in 4℃ （4T group） and group UDP-Gal glycosylated and preserved in 4℃ （ U ＋4T group）. The binding rate of RCA I lectin and expression of platelet surface markers CD62P, CD42b were determined by flow cytometry. Morphology and ultrastructure of platelets were observed by light microscopy, scanning electron microscopy （SEM） and transmission electron microscopy （TEM）. Platelets aggregation was detected by aggregometer. The results showed that the binding rate of RCAI in U ＋4T group significantly higher than that in RT group （p 〈0.01 ）, no obvious changes was found in ultrastruclure of glycosylated platelets, as compared with fresh platelets. Some morphologic changes, such as pseudopodium could be observed in 4T group. The aggregation rate of platelets in U ＋4T group reached to 50% of RT group. The expression levels of CD42b and CD62P, and the binding rate of annexin V in U ＋ 4T group were not significantly different from that in RT group. It is concluded that UDP-Gal can effectively cause galactosylation of platelets, and the platelets modified with UDP-Gal remain normal morphology, ultrastructure and function.
出处
《中国实验血液学杂志》
CAS
CSCD
2008年第2期411-415,共5页
Journal of Experimental Hematology
关键词
血小板
糖基化修饰
超微结构
功能评价
platelet
galactosylated modification
ultrastructure
function assesment
