sRAGE对缺氧/复氧心肌细胞氧化应激的影响

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。

Influence of soluble form of receptor for advanced glycation end products(sRAGE) on hypoxia/reoxygenation myocardial oxidative stress

Objective 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 MTT; 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.