【摘要】 本研究探讨在M-sol(mixture of solutions)血小板保存液中添加花生四烯乙醇胺(N-arachidonoylethanolamine,ANA)对体外保存血小板的影响。利用Amicus血细胞分离机采集无偿献血志愿者的血小板,血小板保养液为M-sol,实验组加入终浓度为0.1-50μmol/L的ANA,未加入ANA的另一组作为对照,置于22±2℃振荡仪中保存。于第7 d取样,用MTT比色法分析血小板的存活率。选定最佳浓度的ANA加入血小板保存液中,分别于第1、5、7、9和11 d取样,检测血小板计数(BPC)、平均血小板体积(MPV)、血小板分布宽度(PDW)、血小板磷脂酰丝氨酸(PS)膜外表达以及可溶性P-选择素(sP-selectin)含量。结果发现,加入终浓度为0.5μmol/L ANA的血小板存活率(91.23±5.44%)最高,与对照组(62.54±4.79%)相比,差异具有统计学意义(P<0.05);保存第1、5、7、9和11 d实验组与对照组BPC均有下降趋势,但两组相比差较异均无统计学意义(P均>0.05);保存期间血小板MPV和PDW有增加趋势,实验组与对照组相比较差异均无统计学意义(P均>0.05)。保存到第9和11 d血小板膜外PS的表达,在实验组PS表达阳性率显著低于对照组(7.69±1.82%vs 11.21±2.03%;10.74±1.78%vs15.37±1.95%),两组相比较差异具有统计学意义(P均<0.05);保存期间可溶性P-选择素含量在两组中均有增加趋势,实验组可溶性P-选择素含量显著低于对照组(30.19±2.03 ng/mL vs 39.18±2.66 ng/mL;34.52±2.64ng/mL vs 43.23±2.58 ng/mL),两组相比差较异具有统计学意义(P均<0.05)。结论:将低浓度ANA加入血小板M-sol保存液中,在一定程度上可以减轻血小板贮存损伤。更多还原

【Abstract】 This study was purposed to investigate the effects of N-Arachidonoylethanolamine(ANA) on the quality of platelets(Plt) stored in Plt M-sol preservative solution at 22±2℃.Samples taken from collecting apheresis Plt by the Amicus instrument and splited into two equal parts were stored in Plt M-sol preservative solution on a shaker at 22±2℃.Different working concentrations of ANA(from 0.1 to 50 μmol/L) were then added into one part of stored Plt as the experimental group,the other without ANA was used as the control group.The viability of Plts stored at 22±2℃ for 7 days was evaluated by MTT colorimetric assay.The most effective concentration of ANA was selected and added to the subsequent experimental group.Plt count(BPC),mean Plt volume(MPV),Plt distribution width(PDW),phosphatidyl serine(PS) and soluble P-selectin were detected on the 1st,5th,7th,9thand 11thday of storage.The results showed that the most effective working concentration of ANA was 0.5 μmol/L,which showed significant increasing Plt viability(91.23 ±5.44%) compared to the control group(62.54 ±4.79%).Thus,ANA concentration at 0.5 μmol/L was chosed to perform subsequent experiments.During 11 days of storage,the BPC,MPV and PDW were not changed significantly between the experimental group and control group,although there was decreasing trend in the BPC and increasing trends in MPV and PDW in the two groups.The rate of Plt PS positive was enhanced during the storage period:the rate of PS positive in experimental group increased from 7.69±1.82% to 10.74±1.78% while it in control group increased from 11.21±2.03% to 15.37±1.95%,with significant differences between the two groups(P<0.05) on the 9thand 11th day of storage,respectively.Soluble P-selectin contents in experimental group on the 9thand 11thday of storage were 30.19±2.03 ng/ml and 34.52±2.64 ng/mL,respectively,while those in control group were 39.18±2.66 ng/ml and 43.23±2.58 ng/ml,respectively,with significant differences between the two groups(P<0.05).It is concluded that the extended storage of Plt in M-sol treated with low concentration ANA can potentially alleviate Plt storage lesions.更多还原