摘要
目的：观察高渗液（HS）对缺氧/复氧的人脐静脉内皮细胞（HUVECs）内游离钙离子浓度（[Ca^2＋]i）的影响,阐明高渗液治疗休克时对血管内皮细胞的保护机制。方法：体外分离和培养的HUVECs分为对照组和实验组,采用荧光探针技术,应用激光共聚焦显微镜,测定细胞内[Ca^2＋]i的变化,两组细胞先置于缺氧（95%氮气,5%CO2）环境中8 h,然后将实验组细胞置于配制好的HS中15 min,再在常氧状态下（95%空气,5%CO2）培养16 h。对照组细胞加对照液（M199培养液）15 min后同实验组。两组细胞分别于缺氧前（T0）、缺氧后（T1）、处理后（T2）、复氧后1/2、2、8、16 h（T36）测定细胞内Fluo-3荧光强度。结果：两组细胞荧光强度T1与T0比较显著升高（P〈0.05）;实验组细胞荧光强度T2低于实验组T1及对照组T2（P〈0.05）,但仍高于T0（P〈0.05）,且复氧过程中逐渐降低,至T5恢复到T0水平;两组细胞T3时点荧光强度均高于各T2时点（P〈0.05）;实验组复氧过程中不同时点（T36）荧光强度低于各自同一时间点的对照组（P〈0.05）。结论：缺氧/复氧可以引发HUVECs内[Ca^2＋]i升高,而HS能减轻HUVECs内的钙超载,保护缺氧/复氧损伤的内皮细胞。
Objective To observe the effects of hypertonic solution（HS） on [Ca^2＋]i in cultured human umbilical vein endothelial cells（HUVECs）suffering from hypoxia/reoxygenation in vitro,and clarify the mechanism of protective effect of HS on vascular endothelial cells（VEC）.Methods HUVECs were successfully isolated by irrigative kneading digestion method,and were seperated to control group and experimental group.HUVECs were cultivated in hypoxia environment（95% nitrogen,5% CO2） for 8 h and placed in HS for 15 min（experimental group）,then cultivated under normal condition（95% air and 5% CO2） for 16 h.HUVECs in control group were placed in M199 for 15 min,then cultivated under normal condition.Fluo-3 fluorescence intensity was detected before experiment（T0）,and in hypoxia（T1）,after treatment（T2）,1/2,2,8,16 h after reoxygenation（T3-6）.Results The fluorescence intensity at T1 was significantly increased compared with T0（P〈0.05）in two groups;in experimental group,the fluorescence intensity at T2 wassignificantly lower than that in control group（P〈0.05）at T2 or experimental group at T1,but still higher than T0（P〈0.05）,and in the process of reoxygenation,the fluorescence intensity gradually reduced to the level of T0 at T5;in the process of reoxygenation,the fluorescence intensities in the experimental group were lower than control group at the same time（T3,T4,T5,T6）（P〈0.05）.Conclusion [Ca^2＋]i in HUVECs increase significantly after hypoxia and reoxygenation.HS can ease the calcium overload in HUVECs,reduce i in HUVECs,then play a protective effect on HUVECs.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2009年第4期612-615,765,F0002,共6页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅科研基金资助课题(200505225)
关键词
人脐静脉
血管内皮细胞
钙离子
高渗液
缺氧/复氧
human umbilical vein
vascular endothelial cells
calcium ion
hypertonic solution
hypoxia/reoxygenation
