辐照对白膜制品中淋巴细胞和粒细胞的影响  

Function of irradiated polymorphonuclear leukocytes and lymphocytes obtained by buffy-coat centrifugation

作　　者：王承琳[1] 张锡敏[1] 崔靖怡 康娜 纪蓉蓉 WANG Chenglin;ZHANG Ximin;CUI Jingyi;KANG Na;JI Rongrong(Tianjin Blood Center,Tianjin 300110,China)

机构地区：[1]天津市血液中心,天津300110

出　　处：《中国输血杂志》2020年第7期654-657,共4页Chinese Journal of Blood Transfusion

摘　　要：目的探讨经辐照处理的白膜制品中的中性粒细胞、淋巴细胞功能的变化。方法随机选取30份健康全血制备白膜制品,将每袋白膜平均分成2组,其中1组接受25 Gy射线辐照即为辐照组,另1组为对照组。分别用裂解液裂解红细胞并将剩余细胞用流式细胞仪分选收集实验细胞。用cck-8检测采集24 h内淋巴细胞增殖;将对照组和辐照组粒细胞分别按采血时间分成6 h组、12 h组、18 h组,每组10份,用流式细胞仪分析各组:FITC标记大肠杆菌检测粒细胞吞噬、活性氧、Annexin V-FITC检测凋亡(早期凋亡、坏死)。结果 1)淋巴细胞辐照前后增殖试验的OD值分别为对照组1.38±0.08,辐照组0.45±0.06(P<0.05);2)辐照前后粒细胞吞噬功能分别为2 630±107,2 390±106(P>0.05)、凋亡分别为5.01±1.16,8.38±1.26(P>0.05)、坏死分别为4.93±1.35,4.89±1.19(P>0.05),但是粒细胞释放的活性氧含量分别为2 829±149,3 373±170(P<0.05);3)吞噬功能:6 h辐照组、12 h辐照组和18 h辐照组分别为2 735±195,2 510±136,1 925±122(6 h、18 h比较,P<0.05;12 h、18 h比较,P<0.05)。活性氧:12 h辐照组、18 h辐照组分别为3 263±290,4 264±378(P<0.05);18 h辐照前后分别为2 932±186,4 264±378(P<0.05)。早期凋亡:6 h辐照前后分别为0.40%±0.26%,1.78%±0.49%(P<0.05);12 h辐照前后分别为3.23%±0.76%,7.66%±1.55%(P<0.05);18 h辐照前后分别为11.41%±2.26%,17.70%±1.94%(P<0.05);6 h、12 h、18 h两两比较都存在显著性差异(P<0.05)。结论 25Gy的辐照有效减少淋巴细胞增殖但会在一定程度影响白膜中的粒细胞功能。临床输注白膜制品应尽早输入,且输注前进行辐照处理。Objective To determine whether polymorphonuclear leukocytes(PMN)and lymphocytes isolated by gamma-irradiated(25 Gy)buffy coat centrifugation have adequate function.Methods Buffy coat centrifugation were prepared from 30 healthy whole blood samples randomly selected.One-half of each buffy coat centrifugation was gamma-irradiated(25 Gy)as the irradiation group,and the other half worked as the control group.After erythrocytes were lysed,the remaining cells were isolated and collected by flow cytometry.Lymphocyte proliferation within 24 h was detected by cck-8.PMNS,isolated either from control group or irradiation group,were divided into 3 groups according to storage time after collection,that is 6 h,12 h and 18 h group(10 samples/group),and each group was analyzed using flow cytometry:oxidative and phagocytic activity were measured by FITC-labeled Escherichia coli.and apoptosis(early apoptosis/necrosis)was measured by Annexin V-FITC.Results 1)The lymphocyte proliferation was effectively prevented with irradiation,as OD value were 1.38±0.08(control group)and 0.45±0.06(irradiation group),respectively(P<0.05).2)For control group and irradiation group,PMNS phagocytic activity was 2630±107 and 2390±106(P>0.05),apoptosis was 5.01±1.16 and 8.38±1.26(P>0.05),necrosis was 4.93±1.35 and 4.89±1.19(P>0.05),more superoxide,however,was released by PMNS from irradiation group significantly(3373±170 vs 2829±149,P<0.05).3)phagocytic activity in 6 h,12 h and 18 h irradiation group was 2735±195,2510±136 and 1925±122,respectively(significant difference was observed between 6 h and 18 h group,12 h and 18 h group,P<0.05).Superoxide release in 12 h and 18 h irradiation group were 3263±290 and 4264±378(P<0.05);in 18 h control group and 18 h irradiation group were 2932±186 and 4264±378(P<0.05).Early Apoptosis(%)in control group and irradiation group at 6 h/12 h/18 h storage was 0.40±0.26 vs 1.78±0.49,3.23±0.76 vs 7.66±1.55,and 11.41±2.26 vs 17.70±1.94,respectively(P<0.05).Conclusion 25 Gy irradiation could effectively

关 键 词：辐照 白膜 增殖 吞噬功能 活性氧 凋亡 

分 类 号：R457.1[医药卫生—治疗学] R331.142[医药卫生—临床医学]