肢体缺血预适应减轻人脐静脉内皮细胞的氧化应激损伤

摘要

AIM: To investigate the effects of the sera from the rats after limb ischemic preconditioning (LIPC) on human umbilical vein endothelial cells (HUVECs) injured by hydrogen peroxide (H2O2).METHODS:The HUVECs were divided into 5 groups:the cells in control group were cultured without any intervention;the cells in model group (M) were damaged by 1 mmol/L H2O2 for 2 h;the cells in early preconditioning serum (EPS) group, delayed pre-conditioning serum (DPS) group or sham limb ischemic preconditioning serum (SPS) group were treated with the corre-sponding serum at 5%for 12 h, respectively , and then treaed with H 2 O2 for 2 h.The viability of the HUVECs was ana-lyzed by flow cytometry .The lactate dehydrogenase ( LDH) in the culture media was detected .The cell adhesion molecules in the HUVECs were detected by real-time PCR.The mRNA and protein expression of heme oxygenase-1 (HO-1) was also determined .RESULTS:The viability of HUVECs incubated with 1 mmol/L H2 O2 for 2 h significantly decreased compared with the control cells , which was accompanied with the augmentations of LDH in the medium and the cell adhesion mole -cules in cells , such as vascular cell adhesion molecule-1 ( VCAM-1 ) and intercellular adhesion molecule-1 ( ICAM-1 ) . Preincubation with EPS and DPS derived from the rats subjected LIPC attenuated these injuries .Furthermore, pretreatment with EPS and DPS increased the expression of HO-1 at mRNA and protein levels .CONCLUSION:LIPC protects the HU-VECs from H2O2-induced injury.%目的：探讨肢体缺血预适应对氧化应激损伤的血管内皮是否具有保护作用。方法：人脐静脉内皮细胞分为5组：对照组不加任何处理；模型组给予过氧化氢损伤2h；早期肢体缺血预适应组、晚期肢体缺血预适应组和假肢体缺血预适组这3组分别用5％大鼠早期肢体缺血预适应血清、晚期肢体缺血预适应血清和假肢体缺血预适应血清孵育12 h，再用过氧化氢损伤2 h。检测细胞凋亡情况，培养液中乳酸脱氢酶的活性，内皮细胞表面黏附分子的mRNA水平，血红素氧合酶1的mRNA和蛋白表达情况。结果：大鼠早期肢体缺血预适应和晚期缺血预适应血清可以减轻人脐静脉内皮细胞氧化应激所导致的细胞死亡，减少乳酸脱氢酶的释放，减少内皮细胞表面黏附分子的表达，并能增加血红素氧合酶1的mRNA和蛋白的表达。结论：肢体缺血预适应可以减轻血管内皮细胞的氧化应激损伤。