摘要
目的观察可溶性晚期糖基化终产物(soluble form of receptor for advanced glycation end products,sRAGE)对缺氧/复氧心肌细胞氧化应激的影响。方法乳鼠心肌细胞原代培养48 h,以低氧3 h、复氧2 h复制缺氧/复氧损伤模型,采用抽签法将乳鼠心肌细胞随机分为4组:常氧对照组(Control,Con)、常氧+sRAGE组(Con-sRAGE)、缺氧/复氧组(Hypoxia/reoxygenation,H/R))、缺氧/复氧+sRAGE组(H/R-sRAGE)。采用四甲基偶氮唑蓝〔3(4,5-dimethyl-thiazol)-2,5-diphenyl-tetrazolium,MTT〕比色法检测心肌细胞活力和培养液中乳酸脱氢酶(lactate dehydrogenase,LDH)浓度,黄嘌呤氧化酶法测定超氧化物歧化酶(superoxide dismutase,SOD)活力,硫代巴比妥酸显色法测定丙二醛(malondialdehyde,MDA)含量,2',7'-二氯荧光黄双乙酸盐(DCFH-DA)荧光探针联合流式细胞仪检测细胞荧光强度-反应活性氧(reactive oxygen species,ROS)水平,硝酸还原酶法测定一氧化氮(nitric oxide,NO)含量。结果与H/R组相比,H/R-sRAGE组可以提高心肌细胞活力,减少LDH漏出量,增加SOD活力,降低MDA、ROS、NO含量(P<0.05)。结论 sRAGE可以直接作用于心肌细胞拮抗缺氧/复氧损伤,其保护性作用与抑制氧化应激有关。
To elucidate the influence of soluble form of receptor for advanced glycation end products （sRAGE） on hypoxia/reoxygenation myocardial oxidative stress. Methods Cardiac myocytes were isolated from neonatal rats with the modified 2-step collagenase digest method and were subjected to Primary culture for 48 hours. Hypoxia 3 h/reoxygenation 2 h injury model was produced. The cells were randomly divided into four groups ： normoxic control group （ Con）, normoxia ＋ sRAGE group （ Con-sRAGE ）, hypoxia/ reoxygenation （H/R） group（ model group） , hypoxia/reoxygenation ＋ sRAGE （H/R-sRAGE） group （ experimental group）. The viability of myocardial cells was detected by M＇I~ ; The leakage of lactate dehydrogenase in culture medium （ LDH ） ; the activity of superoxide dismutase （SOD） were detected by xanthine oxidase; The content of malondialdehyde （MDA） was detected by thiobarbituric acid color method; The intensity of fluorescence was detected by DCFH-DA fluorescent probe combined flow cytometry-Reactive oxygen species （ROS） levels in response; nitrate reductase determination of nitric oxide （NO） levels. Results Compared with H/R group, H/R- sRAGE group could improve the myocardial viability, reduce the amount of LDH leakage, increase SOD activity, lower MDA and ROS levels （P〈0.05）. Conclusion sRAGE may act directly on myocardial cells and antagonize the hypoxia/reoxygenation injury, the protective role is related to inhibition of oxidative stress.
出处
《首都医科大学学报》
CAS
2012年第4期488-493,共6页
Journal of Capital Medical University
基金
国家自然科学基金项目(30801217)
北京市科技新星课题(2010B050)~~
关键词
SRAGE
心肌细胞
缺氧/复氧
氧化应激
soluble form of receptor for advanced glycation end products
myocardial cells
hypoxia/reoxygenation
oxidative stress
