CAJ | 학술논문

目的：探讨贮存期内不同保存时间致悬浮红细胞凋亡损伤及可能的机制。方法采集12例健康献血员血液,常规去除白细胞和血浆。取保存后3、11、19、27、35 d 5个时相点的红细胞,用凋亡试剂染色经流式细胞仪检测红细胞凋亡率及红细胞体积大小变化,用Fluo-3 AM染色经流式细胞仪检测细胞内Ca2+浓度。同时留取上清液,利用酶标仪检测血红蛋白在405 nm的吸光度。结果与保存3 d的红细胞比较,红细胞凋亡率和细胞内Ca2+浓度逐渐增加,至27 d与35 d差异均有统计学意义(P<0．05,P<0．01)；溶血率早期增加不明显,至35 d 差异有统计学意义( P <0．01)；而红细胞体积逐渐变小,至27 d与35 d差异有统计学意义( P<0．05,P<0．01)。结论体外贮存的悬浮红细胞随着保存时间的延长,凋亡逐渐增加,可能是储存损伤的原因之一。Ca2+内流可能是凋亡主要的发生机制。
목적：탐토저존기내불동보존시간치현부홍세포조망손상급가능적궤제。방법채집12례건강헌혈원혈액,상규거제백세포화혈장。취보존후3、11、19、27、35 d 5개시상점적홍세포,용조망시제염색경류식세포의검측홍세포조망솔급홍세포체적대소변화,용Fluo-3 AM염색경류식세포의검측세포내Ca2+농도。동시류취상청액,이용매표의검측혈홍단백재405 nm적흡광도。결과여보존3 d적홍세포비교,홍세포조망솔화세포내Ca2+농도축점증가,지27 d여35 d차이균유통계학의의(P<0．05,P<0．01)；용혈솔조기증가불명현,지35 d 차이유통계학의의( P <0．01)；이홍세포체적축점변소,지27 d여35 d차이유통계학의의( P<0．05,P<0．01)。결론체외저존적현부홍세포수착보존시간적연장,조망축점증가,가능시저존손상적원인지일。Ca2+내류가능시조망주요적발생궤제。
Objective To discuss the effect of different blood storage time on apoptotic lesion of suspended erythro-cytes and possible mechanisms in the preservation period. Methods The blood was collected from 8 healthy volun-teers. After the leukocyte and plasma were depleted, the suspended erythrocytes were preserved as usual. At 5 time points including 3,11,19,27,35 days of blood storage, erythrocyte apoptotic rate and the change of cell volume were measured by Annexin V-FITC apoptosis detection reagent staining and flow cytometry. Fluo 3-AM staining and flow cytometry were used to measure the intracellular free calcium. At the same time the supernatant of the samples was harvested. Using microplate reader, the hemoglobin ( Hb) absorbance of the supernatant was determined at 405 nm. Results Compared with the stored erythrocyte suspensions of 3rd day, erythrocyte apoptotic rate and intra-cellular free calcium were increasing gradually. After 27 days of storage,there was significant difference (P<0. 05, P<0. 01);the increase of hemolytic rate was not obvious in the early days, but there was statistically significant difference on 35 days of storage (P<0. 01);the cell volume was small gradually, but which was remarkably smal-ler on 27 day than that of 3rd day( P<0. 05,P<0. 01). Conclusion With increase in storage time, the increase of erythrocyte apoptotic rate that may be one of the causes of storage lesion. The change of intracellular free calcium may play pivotal roles in apoptosis of erythrocyte.