保存时间对去白悬浮红细胞上清中血小板相关生长因子蓄积的作用及其对肿瘤细胞体外增殖的影响研究  

Effect of Storage Time on Cumulation of Platelet-related Growth Factors in the Supernatant of Leukoreduced Packed Red Cells and Tumor Cell Proliferation In Vitro

作　　者：庄远[1] 张婷[1] 魏超[1] 潘纪春[1] 王淑芳[1] 张爱群[2] 汪德清[1] 

机构地区：[1]中国人民解放军总医院输血科,北京100853 [2]中国人民解放军总医院肝胆外科研究所,北京100853

出　　处：《中国实验血液学杂志》2015年第2期541-545,共5页Journal of Experimental Hematology

基　　金：2010年卫生部行业专项临床有效输血及血液风险控制技术应用研究与推广--子课题"红细胞输注与保存技术的应用研究与推广"(201002005)

摘　　要：目的:探讨保存时间对储存前去白红细胞(LR-pRBC)上清中血小板相关生长因子蓄积的作用及其对肿瘤细胞体外增殖的影响。方法:悬浮红细胞(pRBC)经200 ml红细胞白细胞滤器去除白细胞;随后用四联袋等量分装。所有血袋置于2℃-6℃常规保存。于保存第0、14、21和35天各取1分装袋,以1 006×g离心10 min后留取上清。采用ELISA方法测定血小板衍生生长因子(PDGF)和血管内皮生长因子(VEGF)的蓄积浓度。体外培养Hep G2肝癌肿瘤细胞贴壁后,加入保存第0天和35 d LR-pRBC上清,与Hep G2细胞共同孵育48 h后,应用MTT法测定上清吸光度,以反映肿瘤细胞的体外增殖程度。结果:LR-pRBC上清中VEGF和PDGF浓度蓄积量随保存期延长进一步升高,VEGF在保存末期35 d时,浓度达900.16(95%CI,552.26-1248.07)pg/ml,而0 d LR-pRBC上清中浓度为[611.84(95%CI,356.45-867.23)pg/ml],其浓度有统计学差异(P<0.05)。PDGF在第0天LR-pRBC上清中的初始浓度为2.23(95%CI,0-5.37)ng/L,35 d末其浓度蓄积达5.66(95%CI,-1.16-12.48)(P=0.073)。LR-pRBC上清同Hep G2细胞体外共孵育48 h后,保存35 d的LR-pRBC上清OD值为0.40(95%CI,0.38-0.42),明显促进Hep G2细胞的增殖(P<0.05)。结论:LR-pRBC中残留血小板经2℃-6℃储存后,激活、崩解、释放颗粒物质,造成PDGF和VEGF随保存期延长的进一步蓄积,从而导致保存期末的LR-pRBC上清促进肿瘤细胞体外增殖。Objective：To investigate the effect of storage time on accumulation of platelet-related growth factors in the supematant of leukoreduced packed red blood cells （LR-pRBC） and on tumor cell proliferation in vitro. Methods： LR-pRBC were quartered and stored at 2 ℃ - 6 ℃. The supematant of pRBC was obtained by centrifugation with 1 006 x g for 10 min at day 0, 14, 21 and 35 d. The enzyme-linked immunosorbent assay （ELISA） was used to detect the expression of platelet-derived growth factor（PDGF） and vascular endothelial growth factor （VEGF）. After HepG2 cells was cultured with the supernatant of LR pRBC at day 0 and day 35 together for 48 hours, methylthiazoliltetracolium （MTI＇） method was used to measure the proliferation of tumor cells in vitro. Results： The concentrations of 2 cytokines were still increased with the storage time prolonging. As compared to LR-pRBC at day 0 1611.84 （95% CI, 356.45 - 867.23 ） pg/ml ], the level of VEGF reached 900.16 （95 % CI, 552.26 - 1248. 07 ） pg/ml （ P 〈 0.05 ）. There was a similar tendency in PDGF level with less increment in the supernatant of LR pRBC at day 35 E2.23 （95% CI, 0 - 5. 37） ng/L vs 5.66 （95% CI, 0 -12.48 ） , P = 0.073 ], but there was no significant statistical difference. Likewise, in vitro study of HepG2 cell proliferation showed that the LR-pRBC at day 35 promoted more proliferation of tumor cellswith OD value 0.40 （ 95% CI, 0.38 - 0.42 ） （ P 〈 0.05 ）. Conclusion： The residual platelets in LR-pRBC were activated, disintegrated and released the dense granules and a-granules which induce the accumulation of VEGF and PDGF. It seemed that the supernatant of LR-pRBC promoted the proliferation of tumor cells in vitro.

关 键 词：悬浮红细胞 白细胞去除 保存期 血小板相关生长因子 肿瘤 增殖 

分 类 号：R457.1[医药卫生—治疗学]