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褪黑素对过氧化氢诱导血管内皮细胞损伤的拮抗作用及其机制

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2013年第2期
页码:
158-162
栏目:
基础研究
出版日期:
2013-04-25

文章信息/Info

Title:
Antagonism and mechanism of melatonin against H2O2-induced injury of vascular endothelial cells
作者:
梁振兴1; 杨 阳1; 2; 金振晓1; 王宁1; 易定华1; 段维勋1; 陈文生1
(第四军医大学西京医院心血管外科，陕西 西安 710032；2.北京军区北戴河疗养院医务部医疗科，河北 秦皇岛 066100)
Author(s):
LIANG Zhen-xing1;  YANG Yang1; 2;  JIN Zhen-xiao1;  WANG Ning1;  YI Ding-hua1;  DUAN Wei-xun1;  CHEN Wen-sheng1
(1.Department of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi, China; 2.Department of Medical Administration, Beidaihe Sanatorium, Beijing Military Area Command, Qinhuangdao 066100, Hebei, China)
关键词:
褪黑素; 人脐静脉内皮细胞; 一氧化氮; 乳酸脱氢酶; 过氧化氢
Keywords:
melatonin;  human umbilical vein endothelial cell;  nitric oxide;  lactate dehydrogenase;  hydrogen peroxide
分类号:
R979.9
DOI:
-
文献标识码:
A
摘要:
目的:研究不同浓度褪黑素（melatonin，MLT）对人脐静脉内皮细胞(HUVECs)活力的影响，以及对过氧化氢(H2O2)诱导血管内皮细胞损伤的拮抗作用及机制。方法： 分别用不同浓度（125、250、500 μmol/L）的MLT处理体外培养的HUVECs 2 h、4 h和6 h后，用噻唑蓝（MTT）比色法检测不同浓度的MLT对HUVECs存活率的影响。以300 μmol/L的H2O2建立HUVECs损伤模型，用125、250、500 μmol/L的MLT与H2O2（300 μmol/L）共同处理HUVECs 2 h、4 h和6 h后，分别用噻唑蓝（MTT）比色法测细胞的存活率、细胞黏附实验评价细胞黏附能力、酶活性测定法检测细胞培养液中乳酸脱氢酶(LDH)的释放量和细胞超氧化物歧化酶(SOD)的活力。用Griess法测定细胞培养液中NO-2的浓度来反映HUVECs的一氧化氮(NO)释放量。结果： 以不同浓度的MLT处理内皮细胞 2 h、4 h和6 h后，内皮细胞的存活率无统计学差异。MLT能明显提高H2O2损伤的内皮细胞的存活率、黏附能力及细胞内SOD的活力（P<0.01），增加培养液中NO含量（P<0.01），降低内皮细胞LDH的释放（P<0.01）。以500 μmol/L的MLT效果最明显。结论： 不同浓度的MLT对HUVECs的存活率均无影响，提示MLT药物浓度的安全范围较大。MLT对H2O2诱导HUVECs损伤的拮抗作用可能与其抗氧化能力和促进内皮细胞合成NO有关。
Abstract:
AIM:To study the effect of melatonin (MLT) on normal vascular endothelial cells as well as the antagonism and mechanism of MLT against H2O2-induced injury of vascular endothelial cells. METHODS: Cultured human umbilical vein endothelial cells (HUVECs) were treated with MLT (125, 250 and 500 μmol/L) for 2, 4 and 6 h and the survival rate of HUVECs was detected with thiazole blue (MTT) assay. Injury of HUVECs was induced by H2O2 (300 μmol/L) treatment; serial concentrations of MLT (125, 250 and 500 μmol/L) was applied for 2, 4 and 6 h. The survival rate, adhesion capacity, SOD activity, LDH release and NO-2 concentration of HUVECs were detected. RESULTS: After treatment with MLT (125, 250 and 500 μmol/L) for 2, 4 and 6 h, the survival rate of HUVECs were not significantly altered. The survival rate, adhesion capacity and SOD activity of H2O2-treated HUVECs improved and LDH release was reduced significantly after MLT treatment (P<0.01). In addition, MLT treatment significantly increased the production of NO in cell culture medium compared with H2O2 treatment alone (P<0.01). Compared with the other concentrations observed, MLT concentration of 500 μmol/L produced a more significant antagonistic effect. CONCLUSION: The safe concentration range of MLT is broad because no significant difference is observed in the effects on the survival rate of HUVECs treated with different concentrations of MLT (125, 250 and 500 μmol/L). MLT treatment may antagonize H2O2-induced injury of cultured HUVECs via its antioxidant capacity and promotion of NO synthesis.

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备注/Memo

备注/Memo:
收稿日期：2012-09-20.基金项目：国家十二五学科支撑计划资助（2011BAI11B20）；国家自然科学基金课题资助（81102687和81070198）；陕西省社会发展计划资助（2012JQ4001）；西京医院学科助推计划项目资助（XJZT09M16和XJZT10M11） 通讯作者：陈文生，主任医师，主要从事心血管病的基础研究和外科治疗研究 Email:chenws@fmmu.edu.cn 共同通讯作者：段维勋，主治医师，主要从事抗心肌缺血/再灌注损伤的基础研究 Email: duanweix@fmmu.edu.cn 作者简介：梁振兴，硕士生 Email:liangzhenxing100@163.com
更新日期/Last Update: 2013-04-28