sRAGE对缺氧/复氧心肌细胞氧化应激的影响

doi:10.3969/j.issn.1006-7795.2012.04.015

首都医科大学学报 ›› 2012, Vol. 33 ›› Issue (4): 488-493.doi: 10.3969/j.issn.1006-7795.2012.04.015

sRAGE对缺氧/复氧心肌细胞氧化应激的影响

张敏¹, 郭彩霞¹, 曾翔俊², 王红霞², 张立克², 杜凤和¹

1. 首都医科大学附属北京天坛医院心内科, 北京 100050;2. 首都医科大学基础医学院病理生理教研室, 北京 100069

收稿日期:2012-01-04 修回日期:1900-01-01 出版日期:2012-08-21 发布日期:2012-08-21
通讯作者: 郭彩霞

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

ZHANG Min¹, GUO Cai-xia¹, ZENG Xiang-jun², WANG Hong-xia², ZHANG Li-ke², DU Feng-he¹

1. Department of Cardiology, Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China;2. Department of Pathophysiology, School of Basic medical Sciences, Capital Medical University, Beijing 100069, China

Received:2012-01-04 Revised:1900-01-01 Online:2012-08-21 Published:2012-08-21

摘要/Abstract

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。

关键词: sRAGE, 心肌细胞, 缺氧/复氧, 氧化应激

Abstract: 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.

Key words: soluble form of receptor for advanced glycation end products, myocardial cells, hypoxia/reoxygenation, oxidative stress

中图分类号:

R364.4

张敏;郭彩霞;曾翔俊;王红霞;张立克;杜凤和. sRAGE对缺氧/复氧心肌细胞氧化应激的影响[J]. 首都医科大学学报, 2012, 33(4): 488-493.

ZHANG Min;GUO Cai-xia;ZENG Xiang-jun;WANG Hong-xia;ZHANG Li-ke;DU Feng-he. Influence of soluble form of receptor for advanced glycation end products(sRAGE) on hypoxia/reoxygenation myocardial oxidative stress[J]. Journal of Capital Medical University, 2012, 33(4): 488-493.

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sRAGE对缺氧/复氧心肌细胞氧化应激的影响

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

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