多器官功能障碍综合征患者血清对内皮细胞氧化损伤的影响及还原型谷胱甘肽的保护作用  

Protective effects of GSH against the oxidative damage cultured by MODS serum in vascular endothelial cells

作　　者：张哲英[1] 张蓓[1] 秦静[1] 甄玲玲[1] 马莉[1] 

机构地区：[1]兰州大学第二医院急救中心危重病科,甘肃兰州730030

出　　处：《中国急救医学》2015年第1期38-43,共6页Chinese Journal of Critical Care Medicine

摘　　要：目的探讨多器官功能障碍综合征（multiple organ dysfunction syndrome，MODS）患者血清对人脐静脉内皮细胞（human umbilical vein endothelial cells，HUVEC）氧化损伤的影响及外源性还原型谷胱甘肽（reduced glutathione，GSH）的保护作用。方法选择MODS患者20例，用其血清对HUVEC进行干预，建立体外细胞损伤模型，同时以健康血清组和胎牛血清组作对照，将细胞分为正常对照组（A组）、健康血清组（B组）、MODS血清损伤组（MODS患者m清浓度分别为C1组5％、C2组10％、C3组20％）和GSH干预组（10％MODS患者血清细胞培养基中GSH终浓度分别为D1组0．064mmol／L、D2组0．32mmoL／L、D3组1．6mmol/L、D4组8．0mmol,／L、D5组40mmol／L）。采用MTTr法检测各组细胞生存率；荧光倒置显微镜下观察B组和C2组HUVEC形态学变化；分别检测B组、C2组和D3组药物干预24、48、72h后细胞上清液超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH—Px）活力和总抗氧化能力（T—AOC）水平，同时用流式细胞仪检测细胞内活性氧簇（ROS）水平。结果①B组细胞生存率与A组比较差异无统计学意义（P〉0．05），C1、C2、C3组细胞生存率较B组明显下降（P〈0．05），D2、D3、D4组细胞生存率较C2组明显升高（P〈0．05）；②C2组HUVEC显微镜下可见损伤性改变，细胞变大变圆、界限模糊、胞体碎裂、部分细胞脱落；③C2组细胞培养上清液SOD、CAT、GSH—Px活力及T—AOC水平较B组下降，而D3组各时间点较C2组均明显升高，其中以药物干预72h后升高更明显；④C2组细胞内ROS水平较B组明显升高，D3组各时间点ROS水平较C2组明显降低，其中以药物干预72h后下降最明显。结论MODS患者血清可造成内皮细胞损伤，GSH可通过提高SOD、CAT、GSH—Px活力及T—AOC水平，降低细胞内ROS水平，从而发挥内皮细胞保护作用，减轻血管内皮细�Objective to investigate the oxidative damage in human umbilical vein endothelial cells （HUVECs） cultured in the patient serum with multiple organ dysfunction syndrome （MODS） and the protective effects of reduced glutathione （GSH） on the endothelial dysfunction. Methods MODS serum and control human serum were sampled from twenty MODS patients and twenty healthy controls. HUVECs were cultured in the serum of patients with MODS to establish the injury model. At the same time healthy human serum and fetal bovine serum groups were used as controls. Cells were divided into normal control group （group A）, healthy human serum group （group B）, MODS serum injury group （serum concentration respectively was group C1 ： 5% , group C2 ： 10% , group C3 ： 20% ） and GSH intervention group （ GSH concentration in 10% MODS patient serum separately was group D1 ： O. 064 mmol/L, D2：0.32 mmol/L, D3：1.6 mmol/L, D4： 8.0 mmol/L, D5：40 mmol/L）. The proliferative activity of HUVEC was assessed by blue tetrazolium bromide （MTT）. The cell morphology was observed by inverted microscope in group B and group C2. The supernatant concentrations of superoxide dismutase （SOD）, catalase （CAT）, glutathione peroxidase （GSH- Px） and total antioxidant capacity （T- AOC） were measured in group B, group C2 and group D3 after GSH intervention 24, 48 and 72 h respectively, while intraceUular reactive oxygen species （ROS） level was assessed by flow cytometry. Results There was no significant difference in cell survival rate between group B and group A （P 〉 0.05）. Cell survival rate of group C1, group C2 and group C3 was lower than that in group B （ P 〈 0. 05 ）, while cell survival rate of group D2, group D3 and group D4 was increased higher than that of group C2 （P 〈 0.05 ）. In group C2, the characteristic morphological changes of oxidative injury were observed in HUVECs under transmission microscope. Cells became bigger and round, the boundaries became blurred, cell body fra

关 键 词：多器官功能障碍综合征(MODS) 氧化应激 还原型谷胱甘肽(GSH) 人脐静脉内皮细胞(HUVEC) 

分 类 号：R59[医药卫生—内科学]