张璇之 1龙亚萍 2曹腾宇 2林华雨 2马春娅 2罗圆圆 2刘弈 2孙莉萍 2于洋1
作者信息
- 1. 解放军医学院,北京 100853||中国人民解放军总医院第一医学中心 输血医学科,北京 100853
- 2. 中国人民解放军总医院第一医学中心 输血医学科,北京 100853
- 折叠
摘要
Abstract
Objective To investigate the damaging effects of red blood cell supernatant(RBC-S)stored for different durations(7 d,14 d,and 28 d)on vascular endothelial cells,and to explore the underlying mechanisms using bioinformat-ics analysis,so as to provide references for optimizing red blood cell transfusion strategies.Methods Human umbilical vein endothelial cells(HUVECs)were co-cultured with RBC-S stored for 7,14 and 28 days,designated as the 7 d group,14 d group and 28 d group respectively,which were collectively defined as the experimental groups.Cell damage was evalu-ated by cell proliferation assay(Cell Counting Kit8,CCK8),lactate dehydrogenase(LDH)release assay,4',6diamidino2phenylindole(DAPI)staining,and flow cytometry for apoptosis and reactive oxygen species(ROS)levels.The damage degree of RBC-S on vascular endothelial cells was assessed by statistical analysis of damage data among different groups.Since the damage effect reached a plateau at all time points,the 28 d storage group was selected as the representa-tive for further mechanistic studies.Transcriptomic analysis was performed to explore the role of frizzled class receptor 1(FZD1)and Wnt signaling pathway in red blood cell storagerelated endothelial dysfunction.Results Compared with the control group,the storage groups treated with 7 d,14 d,and 28 d RBC-S showed significantly decreased cell proliferation rates[control group 100%,7 d group(69.51±2.30)%,14 d group(74.54±2.89)%,28 d group(73.59±2.36)%,P<0.05],significantly reduced numbers of DAPI-stained cell nuclei[control group(213±12.5)per field,7 d group(140.33±17.04)per field,14 d group(152.00±23.72)per field,28 d group(144.33±19.09)per field,P<0.05]and significantly increased LDH release[control group(1),7 d group(8.33±1.41),14 d group(9.23±0.83),28 d group(9.16±0.60),P<0.05].There was no significant difference in the degree of damage caused by RBC-S among dif-ferent storage groups(P>0.05).With the prolongation of storage time,free hemoglobin(FHb)gradually increased[con-trol group(not detected),7 d(16.57±6.38)mg/L,14 d(76.80±22.83)mg/L,28 d(286.97±29.02)mg/L,P<0.05].The apoptotic rate(20.53±2.94)%and ROS relative intensity(5.13±0.91)in the 28 d storage group were sig-nificantly higher than those in the control group(P<0.05).Transcriptomic analysis showed that FZD1 played a key role in vascular endothelial dysfunction induced by red blood cell storage and was closely related to the Wnt signaling regulatory net-work.Conclusion RBC-S stored for 7 d,14 d,or 28 d can all significantly damage vascular endothelial cells,and the damaging effect reaches a plateau at 7 d of storage.Mechanistic investigation of the 28 d group indicated that the downregu-lation of the FZD1/Wnt signaling pathway may play a critical role in vascular endothelial dysfunction induced by red blood cell storage,providing a theoretical basis for further optimizing red blood cell storage and transfusion strategies.关键词
红细胞上清液/人脐静脉内皮细胞/储存时间/FZD1/Wnt 信号通路Key words
red blood cell supernatant(RBC-S)/human umbilical vein endothelial cells(HUVECs)/storage time/frizzled class receptor 1(FZD1)/Wnt signaling pathway分类
医药卫生
