储存红细胞与体外传代培养的人脐静脉内皮细胞之间的黏附性研究  

The adhesion effect between the preserved red blood cells and human umbilical vein endothelial cells subcultured in vitro

作　　者：金晶纯[1] 郝一文[1] JIN Jingchun;HAO Yiwen(Department of Transfusion,The First Hospital of China Medical University,Shenyang110001,China)

机构地区：[1]中国医科大学附属第一医院输血科

出　　处：《中国输血杂志》2018年第10期1124-1127,F0001,共5页Chinese Journal of Blood Transfusion

基　　金：沈阳市科技局科学计划项目(17-230-9-53)

摘　　要：目的研究储存红细胞(PRBC)对体外传代培养的人脐静脉内皮细胞(HUVECs)的黏附性。方法载玻片体外传代培养HUVEC细胞株,形成单层正常HUVECs(37℃、5%CO2条件下,RPMI1640培养液培养)及单层损伤HUVECs(0.5%H2O2溶液处理单层正常HUVECs)后,载玻片置于平板流动腔内。将对照组健康人新鲜红细胞(FR-BCs)悬液与实验组储存红细胞(PRBCs)悬液各1mL通过注射器流入并充满平板流动腔。通过细胞低剪切力装置使无血清RPMI1640培养液在连续不同切应力下[1、2、4、6、8、10(dynes/cm2)]流过平板流动腔,通过倒置相差显微镜观察并记录FRBCs与PRBCs分别对正常、损伤HUVECs的黏附数量。结果FRBCs与PRBCs比较:对正常HU-VECs的初始黏附数量(个)为14.1±4.3vs38.2±7.6(P<0.05);对损伤HUVECs的初始黏附数量(个)为28.5±5.6vs76.8±9.7(P<0.05);施加低切应力2、4、6、8、10(dynes/cm2时,对正常HUVECs黏附率(%)分别为12.2±3.9vs25.6±5.3、10.5±3.1vs23.2±5.1、4.3±2.8vs18.8±4.8、2.1±0.7vs10.2±3.3、1.2±0.3vs6.6±2.4(P<0.05);对损伤HUVECs黏附率(%)分别为30.8±6.7vs72.2±8.1、28.3±5.8vs65.6±6.3、22.0±4.4vs56.1±5.5、13.8±3.7vs45.2±4.5,8.6±1.5vs30.7±3.9(P<0.05)。结论无论是FRBC还是PRBCs对正常或损伤HUVECs的黏附率随着切应力升高而降低。损伤HUVECs加重了红细胞黏附,尤其对于PRBCs的黏附有促进作用。Objective To study the adhesion effect between preserved red blood cells and human umbilical vein endothelial cells( HUVECs) subculured in vitro. Method Cover slides were covered by normal single layers of HUVECs( HUVECs strain subcultured with RPMI1640 under 37℃,5%CO2 in vitro) and injured single layer HUVECs( normal HUVECs were treated by 0. 5%H2 O2). Cover slides were then put into flat flow cavity. The control group( fresh red blood cells,FRBCs) and the experiment group( preserved blood cells,PRBCs) were respectively infused into the cover slide by syringe,1 m volume. The flat flow cavity was connected with a cell low shear stress device,and RPMI 1640 solution was forced flowed through the normal HUVECs and injured HUVECs under various shearing stress loads( 1,2,4,6,8,10 dynes/cm2),imitating the human blood flow environment. The number of adhered FRBCs or PRBCs was observed under an invert phase-contrast microscope. The adhesion of FRBCs or PRBCs to normal HUVECs and injured HUVECs were observed by inverted phase contrast microscope. Result The initial adhesion number to injured HUVECs of FRBCs compared with PRBCs: 28. 5± 5. 6 vs76. 8±9. 7( P<0. 05). Under the continuous low shear stress loads( 1,2,4,6,8,10 dynes/cm2),the adhesion number of FRBCs and PRBCs to normal HUVECs cultured in vitro were respectively : 12. 2±3. 9 vs 25. 6±5. 3、10. 5±3. 1 vs 23. 2±5. 1、4. 3±2. 8 vs 18. 8 ± 4. 8、2. 1 ± 0. 7 vs 10. 2 ± 3. 3、1. 2 ± 0. 3 vs 6. 6 ± 2. 4,all P < 0. 05; the adhesion number of FRBCs and PRBCs to injured HUVECs were respectively : 30. 8±6. 7 vs 72. 2 ±8. 1、28. 3±5. 8 vs 65. 6 ±6. 3、22. 0±4. 4 vs 56. 1 ±5. 5、13. 8±3. 7 vs 45. 2±4. 5、8. 6±1. 5 vs 30. 7±3. 9,all P<0. 05. Conclusion FRBCs or PRBCs all adhere to normal HUVECs and injured HUVECs,but the number of adhered cells decreases with increased shear stress loads. The injured HUVECs is in favor of red blood cells adhesion,especially for PRBCs.

关 键 词：红细胞 储存 人脐静脉内皮细胞 黏附 

分 类 号：R457.12[医药卫生—治疗学] R331.141[医药卫生—临床医学]